Xylene

S.R. Clough , in Encyclopedia of Toxicology (Third Edition), 2014

Uses

Xylenes is used as a solvent in the manufacturing of chemicals, agricultural sprays, adhesives and coatings, as an ingredient in aviation fuel and gasoline, and every bit a feedstock in manufacturing various polymers, phthalic anhydride, isophthalic acrid, terephthalic acid, and dimethyl terephthalate. Xylenes are also used equally thinners; solvents in paints, inks, rubbers, gums, resins and lacquers; paint removers, and polyester fibers. A smaller portion of the mixed xylenes produced annually are added to gasoline to amend octane ratings. A typical gasoline contains between 1 and half-dozen% xylene.

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Xylene

Stephen R. Clough , in Encyclopedia of Toxicology (Second Edition), 2005

Toxicokinetics

Xylene is primarily absorbed through the mucous membranes and pulmonary organisation. In experimental subjects, ∼lx% of airborne xylene is absorbed from the lung into the bloodstream. Xylene is as well readily captivated from the gastrointestinal tract and through broken or intact skin. Once absorbed, xylene distributes to many tissues in the body, particularly lipid-rich organs, although this occurs to a lesser extent than for benzene. Chemic amending of xylene occurs in the liver and the lung, where the chemical compound is changed to more water-soluble metabolites (the corresponding o-, yard-, and p-toluic acids and/or methylhippuric acrid) so it can be easily excreted in the urine. In animals, it has been shown that metabolism is qualitatively different in the lung versus the liver. Greater than 95% of absorbed xylene is excreted every bit a water-soluble metabolite, with the remaining fraction being exhaled unchanged. Excretion appears to occur rapidly; fauna studies indicate consummate clearance of the compound in 24   h. Xylene will cantankerous the placenta and enter fetal tissue.

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Anti-inflammatory and Anti-nociceptive Activities of African Medicinal Spices and Vegetables

J.P. Dzoyem , ... U. Bakowsky , in Medicinal Spices and Vegetables from Africa, 2017

2.i.1.2 Xylene-induced ear edema assay

Xylene, xylol, or dimethylbenzene is known to cause astringent vasodilation and edematous changes of skin as signs of astute inflammation ( Tang et al., 1984). The increased thickness of ear tissues is caused by these histopathological changes. After topical application of xylene, severe vasodilation and edematous changes of the peel occur, and infiltration of inflammatory cells is detected, providing signs of acute inflammation. Suppression of this response is taken equally a hallmark of antiphlogistic effect (Atta and Alkofahi, 1998). In the screening of spices for antiinflammatory properties, inhibition of the xylene-induced increase in ear weight is regarded every bit prove of antiinflammatory efficacy through reducing vasodilation and hence improving the edematous condition (Okoli et al., 2007; Ishola et al., 2011, 2012, 2013; Ibironke and Odewole, 2012; Sowemimo et al., 2013, 2015)

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Benzene and Related Aromatic Hydrocarbons

DANA B. MIRKIN Medico , in Haddad and Winchester'due south Clinical Management of Poisoning and Drug Overdose (Fourth Edition), 2007

XYLENE

Xylene (dimethylbenzene) is a commonly used effluvious solvent with three isomeric forms: ortho, meta-, and para-xylene. Xylenes are i of the highest-book chemicals produced and used by industry. A mixture of all three isomers is termed xylol. 1 Information technology is a colorless, flammable, liquid hydrocarbon with a characteristic aromatic odor. It is commonly plant in products such every bit varnish, ink, paint thinners, degreasers, and insecticides. 46 Xylenes like benzene and toluene are major components of gasoline and fuel oil. The primary uses of xylenes industrially are as solvents and synthetic intermediates. 15 Commercial xylene is often contaminated with other organic compounds such every bit ethylbenzene, toluene, benzene, trimethylbenzene, phenol, thiophene, and pyridine. The volumes of these contaminants are very minor, making up less than a fraction of 1%.

The scent threshold of xylenes depends on the isomer. On boilerplate, the odor threshold appears to be about 1.0ppm. m-Xylene has an airborne aroma threshold of 3.7ppm; o-xylene, 0.17 ppm; and p-xylene, 0.47 ppm.

Liquid xylene is irritating to the skin and eyes, causing local vasodilation past the liberation of histamine and 5-hydroxytryptamine. Direct xylene eye splash causes initial transient discomfort and hyperemia of the conjunctiva. The cornea may rapidly develop a visually significant central haze usually express to the anterior portions of the stroma. A characteristic clinical feature reveals articulate vacuoles resembling microcystic corneal epithelial edema but localized generally in the inductive stroma. The stromal vacuoles are very characteristic and only mentioned in a few other entities such every bit n-butanol and nitronaphthalene exposure. In contrast, epithelial cystic changes differ from stromal vacuoles and are ordinarily seen in other keratopathies such equally contact lens toxicity and increased intraocular pressure.

The CNS mechanism of toxicity is identical to that of toluene and other hydrocarbons. 36

The toxicokinetics (TK) and acute toxicity of toluene, xylenes, and other effluvious solvents are quite like. Xylenes and the others are well absorbed from the lungs and alimentary canal, distributed to tissues co-ordinate to tissue blood period and lipid content, exhaled to some extent, well metabolized past hepatic P-450s, and largely excreted equally urinary metabolites.

Dermal xylene absorption is low compared with respiratory tract absorption.

Xylene is metabolized through the P-450 mixed-function microsomal enzyme organisation in the endoplasmic reticulum of the liver. The biotransformation of xylene through side-chain oxidation and effluvious oxidation results in metabolites of methylbenzyl alcohols, methylbenzaldehyde, and methylbenzoic acids (toluic acids). Methylbenzoic acids are conjugated with glycine to grade methyl hippuric acids, the main urinary metabolites of xylene. A minor (ane%–4%) metabolic pathway of xylene metabolism is aromatic ring hydroxylation, which forms xylenol.

The major route of clearance is through the kidneys, with metabolites of xylene beingness excreted past a slow phase and a rapid phase. In humans, about 36% of xylene is excreted past the terminate of the daily working period, and about lxx% to fourscore% of metabolites are excreted within 24 hours of cessation of exposure. Removal from fat occurs slowly over a menses of days. 47

The coingestion of ethanol will inhibit the metabolism of xylene through the pathway leading to methylhippuric acrid, just not the path leading to 2,iv-xylenol.

Coexposure to other solvents affects xylene metabolism. Exposure to both methyl ethyl ketone and xylene results in about a 50% increment in claret xylene concentrations. As well, urinary excretion of methyl hippuric acid decreases, indicating competition for enzyme metabolism. Coexposure to trichloroethylene, ethylbenzene, and toluene too inhibits xylene metabolism. Increased use of the minor aromatic ring hydroxylation pathway occurs when other solvents compete with CYP450 enzyme pathway metabolism.

Xylene is irritating to the eyes, skin, and mucous membranes. Exposure may crusade dyspnea, anorexia, nausea, vomiting, and dermatitis. At that place are several ways in which xylene can exist toxic, such every bit through inhalation, ingestion, or direct contact with the liquid. Ocular injuries such as irritation or conjunctivitis have been reported later exposure to vapors or accidental splashes. Boosted ocular toxic changes may include vacuolar keratopathy, which has been reported in simply a few cases with prolonged exposure to loftier vapor concentrations or to mixed solvents.

Attributable to its high vapor force per unit area, most exposures to xylene and its isomers are by inhalation.

Acute CNS effects are identical to those of toluene and other volatile hydrocarbons. Workers rarely progress to the level of intoxication considering the respiratory irritability and odor threshold limit of xylene are such that workers are unlikely to tolerate high exposures necessary to produce these effects. 49 Nevertheless, beast studies lend support to the theory that mixed xylene isomers are neurotoxic later inhalation.

Increased theta waves over occipital regions are seen in electroencephalogram results in subjects exposed to xylene peaks of 200 ppm for 4 hours.

Xylenes appear to accept very limited capacity to adversely impact organs other than the CNS. Mild, transient liver or kidney toxicity has occasionally been reported in humans exposed to high vapor concentrations of xylenes.

Xylene does not appear to exist genotoxic or carcinogenic.

For day-long working conditions, almost experimentally exposed subjects have selected 100 ppm every bit the highest tolerable concentration, with considerable eye irritation in some individuals exposed to 200 ppm in air. Nose and throat irritation from xylene has been reported at 200ppm for iii to 5 minutes and 100 ppm for one to 7.5 hours per solar day for 5 days. Chronic occupational exposure to unspecified or unknown concentrations of xylene vapors has been associated with difficulty breathing and dumb pulmonary office. Nose and throat irritation has been reported with increased prevalence past workers who are exposed chronically to xylene vapors at a geometric mean TWA concentration of 14 ppm.

Expiry was reported in an individual exposed to paint solvents containing primarily xylene at an estimated atmospheric concentration of x,000 ppm. Autopsy demonstrated severe pulmonary congestion with hemorrhage and pulmonary edema.

Liver necrosis and steatosis have been reported after xylene exposure.

Xylene vapors have a sweet scent that, in conjunction with irritation of the airways and respiratory tract, will cause most individuals to avoid exposure to the compound at high concentrations. Those who may be tolerant of the odor or who remain in the area of airborne xylene may develop headaches, nausea, vomiting, fatigue, dizziness, irritability, insomnia, a drunken feeling, impaired retentiveness, loss of coordination, and unsteady gait, in addition to upper airway and ocular irritation.

Like toluene, xylene can also sensitize the myocardium to the arrhythmogenic furnishings of catecholamines.

Assays of blood and alveolar air tin can detect xylene, and its metabolites tin be detected in urine. Claret levels of xylene can be affected by coexposure to other solvents. Coexposure to toluene, trichloroethylene, ethylbenzene, methyl ethyl ketone, or booze is known to increase xylene claret concentrations by competing for metabolic enzymes. Aspirin decreases methylhippuric acid urinary excretion.

Methylhippuric acrid is non normally nowadays in the urine, and methylhippuric acid concentration has, therefore, been proposed every bit a marking for the biologic monitoring of workers exposed to xylene. In full general, methylhippuric acrid levels appear to correlate linearly and significantly with the TWA exposure if urine is collected at the end of the daily working period and, preferably, in the latter half of the workweek when urinary metabolites are probable to attain maximum levels. 47

The BEI for xylene is i.five one thousand methylhippuric acrid per gram of creatinine in urine nerveless at the stop of the work shift. 48

See Box 94-ane for acute clinical management.

After an acute splash in the eye, immediate and thorough irrigation with Ringer's lactate solution or saline is recommended. Delayed irrigation, especially in the presence of normal ocular pH, does non seem to be indicated. Removal of particulate chemical matter from ocular surfaces and conjunctival fornices is crucial. If meaning corneal edema is present, topical steroids may shorten the class. Xylene keratopathy should be considered in the differential diagnosis of epithelial and stromal vacuolar keratopathy.

In acute symptomatic exposures, xylene may be detectable in blood drawn with a gas-tight syringe, simply usually only for a few hours. The metabolite methylhippuric acid is excreted in the urine and can exist used to certificate exposure, but urine levels exercise non correlate with systemic effects.

There is no antidote. At that place is no role for enhanced elimination. Disposition is the same as for toluene.

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Nervous System and Behavioral Toxicology

B. Pouyatos , ... L. Linda , in Comprehensive Toxicology, 2010

13.xiv.7.2.1.(iv) Xylenes

Xylene ( Figure 8a ) is used as a solvent in the press, rubber, and leather industries and is i of the top thirty chemicals produced in the The states. Xylene exists as three different isomers: meta-, ortho-, and para-xylene. Generally, the industrial xylene is used as a blend of those 3 isomers, normally referred to as mixed xylene.

While the neurotoxic effects of xylene in human and animal models have been studied extensively, the literature concerning its ototoxicity is quite limited. Pryor et al. (1987) and Crofton et al. (1994) showed a mid-frequency hearing loss after exposure to mixed xylene by inhalation for half dozen weeks to 800   ppm and 5 days at 1800   ppm, respectively. Both investigations determined that at equal doses xylene was less ototoxic than styrene but more ototoxic than toluene. A few years subsequently Gagnaire et al. (2001) demonstrated that only para-xylene induced severe hearing loss and outer hair cell loss for concentrations of 900   ppm or more (for xiii weeks). Meta- and ortho-xylene did not induce any hearing loss even at the highest dose ( Effigy 13 ). More recently, Maguin et al. (2006) confirmed this result with an exposure to 1800   ppm for 3 weeks, and observed that para-xylene's histopathological signature was similar to toluene's and styrene'southward, that is, injury of the outer hair cells in the middle plow of the organ of Corti.

Effigy 13. Permanent auditory threshold shifts measured in animals exposed to para-xylene, ortho-xylene, and meta-xylene. The solvent exposure level was identical regardless of the isomer (1800 ppm, 6   h solar day−1, five days week−1 for four weeks). Reproduced from Maguin, Thousand.; Lataye, R.; Campo, P.; Cossec, B.; Burgart, M.; Waniusiow, D. Neurotoxicol. Teratol. 2006, 28, 648–656.

The literature contains no human data on occupational exposure to pure xylene. Even so, two contempo epidemiological studies conducted in Poland on workers in pigment and varnish production plants showed that exposure to low doses of solvent mixtures, including mixed xylene, significantly increased the risk of developing hearing loss (Sliwinska-Kowalska et al. 2001; Sulkowski et al. 2002).

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Tissue processing

Lena T. Spencer , John D. Bancroft , in Bancroft's Theory and Do of Histological Techniques (Seventh Edition), 2013

Clearing agents suitable for routine employ

Xylene

Xylene is a flammable, colorless liquid with a characteristic petroleum or aromatic odor, which is miscible with most organic solvents and alkane series wax. Information technology is suitable for clearing blocks that are less than 5 mm in thickness and quickly replaces alcohol from the tissue. Overexposure to xylene during processing can cause hardening of tissues. It is virtually commonly used in routine histology laboratories and is likewise recyclable.

Toluene

This has like backdrop to xylene, although it is less damaging with prolonged immersion of tissue. It is more flammable and volatile than xylene.

Chloroform

Chloroform is slower in action than xylene just causes less brittleness. Thicker tissue blocks can be processed, greater than 1 mm in thickness. Tissues placed in chloroform practise not get translucent. It is not-combustible but highly toxic, and produces highly toxic phosgene gas when heated. It is most commonly used when processing specimens of the central nervous system.

Xylene substitutes

Xylene substitutes are aliphatic hydrocarbons that be in long- and short-chained forms. They differ in the number of carbon atoms inside the carbon chain. Curt-chained aliphatics have the same evaporation properties as xylene, and have no analogousness for water. Long-chained aliphatics practise not evaporate rapidly and may cause contamination of the paraffin wax on tissue processors.

Citrus fruit oils – limonene reagents

Limonene reagents are extracts from orangish and lemon rinds; they are not-toxic and miscible with water. Disposal is dependent upon the water handling centers and local/national standards. The principal disadvantages are that they can crusade sensitization and have a stiff pungent olfactory property that may crusade headaches. Also, small mineral deposits such as copper or calcium may deliquesce and leach from tissues. They are extremely oily and cannot be recycled.

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Membrane Operations in Molecular Separations

T. Uragami , in Comprehensive Membrane Scientific discipline and Engineering science, 2010

ii.11.6.half dozen Selective Membranes for Isomers

Propanol and xylene-isomer-selective membranes are very important for the separation of industrial chemical products and energy saving for the separation of those mixtures.

PVA membranes containing β-CD (PVA/CD membrane) were prepared and the permeation and separation characteristics for propanol isomers through the PVA/CD membranes were investigated by PV and EV [215]. EV was more constructive for the separation of propanol isomers through the PVA/CD membrane than PV. The PVA/CD membrane preferentially permeated 1-propanol rather than 2-propanol from their mixtures. In particular, a mixture of 10 wt.% ane-propanol concentration was full-bodied to nigh 45 wt.% through the PVA/CD membrane. The permeation mechanism of propanol isomers through the PVA/CD membrane was discussed based on the solution–diffusion model.

The characteristics of permeation and separation for xylene isomers through PVA membranes containing β-CD (PVA/CD membranes) were investigated by PV and EV. EV was more effective for the separation of xylene isomers through the PVA/CD membrane than PV. The increase in CD content gave an increase in p-xylene/o-xylene selectivity through the PVA/CD membrane by EV. This was attributed to the stronger affinity of CD for p-xylene compared with o-xylene. Peculiarly, the PVA/CD membrane at a CD content of 40 wt.% showed a higher separation gene for p-xylene/o-xylene selectivity than has e'er been reported. When the p-xylene concentration in the feed was lower, the p-xylene/o-xylene selectivity was improved. The mechanism of permeation and separation for xylene isomers was discussed from the standpoint of solution–improvidence model [216].

PAA membranes containing α-, β-, or γ-CD were prepared and used for the separation of o-/p-xylene mixture by PV. The native PAA membrane was almost impermeable for the xylene isomers, and the incorporation of CDs in the PAA membranes resulted in membranes having molecular recognition function, which selectively facilitated the transport of the xylene isomers. For all types of CDs, the facilitated transport occurred at CD concentrations higher than the threshold concentration. As the CD concentration increased, the permeation rates increased, while the o-xylene/p-xylene selectivities were almost abiding. The selectivity for o-xylene/p-xylene selectivity of the membranes was strongly influenced by the types of CDs incorporated in the membranes [217].

Polymer membranes containing α-CD were prepared by the casting method using cross-linking reaction with hexamethylenediisocyanate. The movie synthesis conducted with and without dibutyltin dilaurate as catalyst resulted in two series of materials in which α-CD host entities were chemically linked to PVA and physically entrapped in PVA, respectively. The obtained membranes were successfully applied to the separation of o-/p- and o-/m-xylene isomer mixtures by pertraction from water. p- and m-xylenes were found to be the faster permeants compared to the o-isomer. The separation factor for p-xylene/o-xylene selectivity varied from 7.75 to 0.35, depending on the membrane α-CD content and the feed concentration. The permeation rate and separation selectivity data were discussed in terms of molecular recognition past α-CD and of coupling ship effect [218].

The PV characteristics of xylene isomer mixtures through a fixed carrier membrane consisting of CA as a base of operations polymer and DNP group equally a selective fixed carrier were studied. In the PV of xylene isomer mixtures, the DNP group selectively facilitated the transport of xylene isomers through the membrane. The order of the preferentially permeating component was p-xylene   > chiliad-xylene   > o-xylene [219].

The selective extraction of the geometrical isomers of xylene using a hydrogel consisting of a PVA grafted with α-CD and β-CD as the complexing moieties was described. The membrane contactors were prepared past the casting method following cantankerous-linking reaction with hexamethylene diisocyanate (HMDI). The transfer of xylenes across the CD-containing membranes was facilitated compared to PVA. The better discrimination was observed for membranes based on α-CD, the more efficient being that containing ∼21 wt.% CD. The gild of permeation rate p-xylene   > m-xylene   > o-xylene followed the affinity social club inferred from the stability constants α-CD. The permeate limerick was independent of the feed composition [220].

Fractionation of o- and p-xylene isomeric mixtures was performed using PV with PU membranes containing ZSM zeolite. The xylene vapor sorption isotherms exhibited a Henry's law relationship in this PU–zeolite blend. In binary solutions, the individual xylene uptake was also proportional to the solvent composition. Although incorporating zeolite into the PU–zeolite membrane rendered a decrease in xylene solubility as compared with that sorbed in the PU film without zeolite addition, the increase of diffusion coefficient and diffusivity selectivity increases enhanced the separation efficiency using the PU-zeolite blend. Increasing the operating temperatures enhanced the xylene permeation rate of xylene. The permeation rates of xylene and selectivity increased with increasing zeolite content [221].

Palygorskite–polyacrylamide (PGS/PAM) hybrid materials were synthesized via intercalation polymerization initiated by redox initiator consisting of modified PGS (reducer) and ceric salt Ce4+ (oxidant), and used as PV membranes. The swelling behavior of hybrid membranes was investigated in single xylene isomer (p-xylene, 1000-xylene, and o-xylene), binary xylene isomer mixtures (p-/o-xylene (the mixtures of p-and o-xylene), o-/g-xylene (the mixtures of o- and m-xylene), and p-/m-xylene (the mixtures of p- and m-xylene)), and ternary isomer mixture (p-/chiliad-/o-xylene (the mixtures of p-, m-, and o-xylene)). A maximum value of caste of swelling at equilibrium (DSequilibrium) in single xylene isomer at xxx   °C exhibited for the hybrid membrane with a PGS of 1.92 wt.%. Negative deviation and both negative and positive deviations of the DSequilibrium based on the improver rule existed in binary or ternary xylene isomer mixtures, respectively, for hybrid membranes with different PGS content at 30   °C. In add-on, a maximum value of separation factor of the hybrid membrane revealed for each pair of binary xylene isomer mixtures when the PGS content was i.92 wt.% in the hybrid membrane. A reversion of the preferential selectivity and a high permeation activation energy of the hybrid membrane occurred at the concentration region with high p-xylene content in p-/1000-xylene binary xylene isomer mixtures. The swelling behavior and PV performance of PGS–PAM hybrid membranes were discussed in terms of the entrapping channel in the PGS for the potential introduction of xylene isomers, the interaction between xylene isomers in feed and the solution–improvidence mechanism in the PV process [222].

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G Protein Pathways, Part B: Grand Proteins and their Regulators

Wei He , Theodore Grand. Wensel , in Methods in Enzymology, 2002

Solutions and Materials

Xylene

Ethanol

Phosphate-buffered saline (PBS) buffer (GIBCO, Grand Isle, NY)

Proteinase K (GIBCO)

NTE: 100   chiliadGrand Tris, pH   seven.six, i   one thousandM EDTA

PFA buffer: 4% Paraformaldehyde in PBS

NTE: 100   mChiliad Tris, pH   8.0, 0.5 M NaCl, ane   mM EDTA

Acetylation buffer: 240 μl acetic anhydride in 100   ml triethanolamine, pH   8.0

SSC: thirty   grandThou Sodium citrate, pH   7.0, 300   mM NaCl

Prehybridization solution:

Formamide 5.0   ml
SSC (20   ×) 2.0   ml
Denhardt's reagent (l   ×) 0.ii   ml
Herring sperm Deoxyribonucleic acid(10   mg/ml) 0.5   ml
Yeast tRNA (10   mg/ml) 0.25   ml
Dextran sulfate (50%) ii.0   ml

Buffer 1: 100   mM Tris, pH   seven.5, 150   mYard NaCl

Buffer 2: 0.3% Triton 10-100 in buffer i

Buffer 3: 100   grandOne thousand Tris, pH   nine.5, 100   m1000 NaCl, 50   mChiliad MgCl2

NBT/BCIP: 50 μl NBT [100   mg/ml in lxx% dimethylformamide (DMF), Roche] and 37.v μl BCIP (50   mg/ml in DMF) and ane   mChiliad levamisole (Sigma, St. Louis, MO) in buffer 3

Coplin jars

Hot shaker (Bellco)

37° water bathroom

42° incubator

Wet sleeping room

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Laser Microdissection

Diane L. Sterchi , in Theory and Exercise of Histological Techniques (Sixth Edition), 2008

Methane series hematoxylin and eosin (H&Eastward) staining method

Slides are deparaffinized using xylene. Xylene substitutes are not platonic to utilise on slides for laser microdissection. Xylene substitutes leave an oily residue that sometimes interferes with the attachment of the cells to the thermoplastic picture show used with ane of the laser microdissection instruments. The following H&Due east staining method is recommended. When looking at protein characterization, continue the hematoxylin staining time to a minimum and omit the eosin.

1.

Xylene × three

 5–8 min (for obviously drinking glass slides)
 Xylene × 2 2–3 min (for membrane/foil slides)
2.

100% ethanol × 2

 1–3 min each
iii.

95% ethanol

 i–xv dips
4.

70% ethanol

 ane–15 dips
five.

Rinse slides briefly in RNase-gratuitous water
6.

Hematoxylin

 thirty–45 seconds.

Use a progressive hematoxylin to avoid the differentiation and bluing steps.

7.

Rinse slides in warm RNase-free water to bluish the hematoxylin.
8.

Eosin

 3–5 quick dips
ix.

95% ethanol

 ane–viii dips
ten. 100% ethanol × two 1–iii min each
11.

100% xylene × 2

 ane–three min each.

After the staining procedure is completed, permit slides to dry out in a cool dust-free area by placing into a slide box or a desiccator with a moisture-arresting material, recalling that moisture encourages RNase activity.

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Mitochondria, 3rd Edition

Sandra Franco-Iborra , Kurenai Tanji , in Methods in Jail cell Biology, 2020

2.ii.two.3 Reagents

▪

Xylene (Sigma Aldrich, Ref. 214736)

▪

Ethanol (Decon Labs, Inc., Ref. 2701)

▪

Distilled water

▪

Hydrogen peroxide (Sigma Aldrich, Ref. 216763)

▪

Phosphate buffer (PBS)

▪

Normal caprine animal serum (Sigma Aldrich, Ref. NS02L)

▪

ABC standard kit (Thermo Fisher, Ref. 32020)

▪

Primary antibodies

○

Anti-complex 3 FeS—gift from Dr. Gonzalez-Halphen (Gonzalez-Halphen, Lindorfer, & Capaldi, 1988)

○

Anti-complex IV subunit Ii—gift from Dr. R. Doolittle (Mariottini et al., 1983)

TIP: For a list of more commercially available antibodies to detect mitochondrial proteins encounter Table i.

▪

Secondary antibodies

○

Goat anti-rabbit HRP secondary antibody (Abcam, Ref. ab205718)

▪

DAB kit (Pierce, Thermo Fisher, Ref. 34002)

▪

Hematoxylin (Sigma Aldrich, Ref. H3136)

▪

DPX mounting medium (Sigma Aldrich, Ref. 06522)

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