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Product Info Summary
| SKU: | RP1111 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | IF, IHC, ICC, WB |
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Product info
Product Name
Anti-Alcohol dehydrogenase class-3 ADH5 Antibody Picoband®
SKU/Catalog Number
RP1111
PB1086 is an alternative SKU for this antibody, used in previous lots.
Size
100 μg/vial
Form
Lyophilized
Description
ADH5 (alcohol dehydrogenase class-3) metabolizes diverse substrates and has high activity for oxidation of S-hydroxymethyl-glutathione (a formaldehyde–glutathione adduct), supporting cellular formaldehyde detoxification and broader aldehyde/alcohol metabolism biology. Assay context: antibody validated for IF, IHC, ICC, and WB; observed molecular weight ~39 kDa is reported on-page, supporting expected-size confirmation in immunoblotting. Often contextualized with aldehyde dehydrogenase pathways such as ALDH3A2 and lysine-catabolism–linked ALDH7A1, or reductase modules like AKR1B1 when mapping oxidative/aldehyde-stress handling across tissues (putative).
Storage & Handling
Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
Cite This Product
Anti-Alcohol dehydrogenase class-3 ADH5 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # RP1111)
Host
Rabbit
Contents
Each vial contains antibody formulated with stabilizing components, 0.9 mg NaCl, 0.2 mg Na2HPO4, and 0.05 mg NaN3.
*This antibody is supplied in a stabilized formulation.
Compatibility with conjugation reactions depends on the chemistry of the conjugation method used.
For conjugation methods that are not compatible with the stabilizing components present in this formulation, a carrier-free antibody format is required.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E. coli-derived human ADH5 recombinant protein (Position: K212-I374). Human ADH5 shares 90.2% and 92% amino acid (aa) sequence identity with mouse and rat ADH5, respectively.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
RP1111 is reactive to ADH5 in Human, Mouse, Rat
Observed Molecular Weight
39 kDa
Calculated molecular weight
39.7 kDa
Background of ADH5
Alcohol dehydrogenase class-3 is an enzyme that in humans is encoded by the ADH5 gene. This gene encodes a member of the alcohol dehydrogenase family. Members of this family metabolize a wide variety of substrates, including ethanol, retinol, other aliphatic alcohols, hydroxysteroids, and lipid peroxidation products. The encoded protein forms a homodimer. It has virtually no activity for ethanol oxidation, but exhibits high activity for oxidation of long-chain primary alcohols and for oxidation of S-hydroxymethyl-glutathione, a spontaneous adduct between formaldehyde and glutathione. This enzyme is an important component of cellular metabolism for the elimination of formaldehyde, a potent irritant and sensitizing agent that causes lacrymation, rhinitis, pharyngitis, and contact dermatitis. The human genome contains several non-transcribed pseudogenes related to this gene.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
RP1111 is guaranteed for IF, IHC, ICC, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.1-0.5μg/ml, Human, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human
Immunocytochemistry/Immunofluorescence, 2μg/ml, Human
Positive Control
WB: human HepG2 whole cell, human K562 whole cell, human MOLT4 whole cell, rat liver tissue, mouse liver tissue
IHC: human liver cancer tissue, human mammary cancer tissue
ICC/IF: A431 cell
Validation Images & Assay Conditions
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Western blot analysis of ADH5 using anti-ADH5 antibody (RP1111).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human HepG2 whole cell lysates,
Lane 2: human K562 whole cell lysates,
Lane 3: human MOLT4 whole cell lysates,
Lane 4: rat liver tissue lysates,
Lane 5: mouse liver tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-ADH5 antigen affinity purified polyclonal antibody (Catalog # RP1111) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for ADH5 at approximately 39 kDa. The expected band size for ADH5 is at 40 kDa.
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IHC analysis of ADH5 using anti-ADH5 antibody (RP1111).
ADH5 was detected in paraffin-embedded section of human liver cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-ADH5 Antibody (RP1111) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
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IHC analysis of ADH5 using anti-ADH5 antibody (RP1111).
ADH5 was detected in paraffin-embedded section of human mammary cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-ADH5 Antibody (RP1111) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
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IF analysis of ADH5 using anti-ADH5 antibody (RP1111).
ADH5 was detected in immunocytochemical section of A431 cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL rabbit anti-ADH5 Antibody (RP1111) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Specific Publications For Anti-Alcohol dehydrogenase class-3 ADH5 Antibody Picoband® (RP1111)
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4 Customer Q&As for Anti-Alcohol dehydrogenase class-3 ADH5 Antibody Picoband®
Question
We are currently using anti-ADH5 antibody RP1111 for human tissue, and we are satisfied with the IHC-P results. The species of reactivity given in the datasheet says human, mouse, rat. Is it likely that the antibody can work on canine tissues as well?
Verified Customer
Verified customer
Asked: 2020-04-24
Answer
The anti-ADH5 antibody (RP1111) has not been validated for cross reactivity specifically with canine tissues, but there is a good chance of cross reactivity. We have an innovator award program that if you test this antibody and show it works in canine you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2020-04-24
Question
Do you have a BSA free version of anti-ADH5 antibody RP1111 available?
Verified Customer
Verified customer
Asked: 2019-05-30
Answer
Thanks for your recent telephone inquiry. I can confirm that some lots of this anti-ADH5 antibody RP1111 are BSA free. For now, these lots are available and we can make a BSA free formula for you free of charge. It will take 3 extra days to prepare. If you require this antibody BSA free again in future, please do not hesitate to contact me and I will be pleased to check which lots we have in stock that are BSA free.
Boster Scientific Support
Answered: 2019-05-30
Question
I was wanting to use your anti-ADH5 antibody for IHC-P for rat lower esophagus on frozen tissues, but I want to know if it has been tested for this particular application. Has this antibody been tested and is this antibody a good choice for rat lower esophagus identification?
Verified Customer
Verified customer
Asked: 2018-06-12
Answer
As indicated on the product datasheet, RP1111 anti-ADH5 antibody has been tested for IF, IHC-P, ICC, WB on human, mouse, rat tissues. We have an innovator award program that if you test this antibody and show it works in rat lower esophagus in IHC-frozen, you can get your next antibody for free.
Boster Scientific Support
Answered: 2018-06-12
Question
See attached the WB image, lot number and protocol we used for lower esophagus using anti-ADH5 antibody RP1111. Please let me know if you require anything else.
R. Jones
Verified customer
Asked: 2017-05-15
Answer
Thank you very much for the data. Our lab team are working to resolve this as quickly as possible, and we appreciate your patience and understanding! You have provided everything we needed. Please let me know if there is anything you need in the meantime.
Boster Scientific Support
Answered: 2017-05-15
