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Product Info Summary
| SKU: | PB9328 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Monkey, Mouse, Rat |
| Host: | Rabbit |
| Application: | IF, IHC, ICC, WB |
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Product info
Product Name
Anti-ATP5H Antibody Picoband®
SKU/Catalog Number
PB9328
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-ATP5H Antibody Picoband® catalog # PB9328. Tested in IF, IHC, ICC, WB applications. This antibody reacts with Human, Monkey, Mouse, Rat. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
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-ATP5H Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9328)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human ATP5H recombinant protein (Position: A2-L161). Human ATP5H shares 81% and 78% amino acid (aa) sequence identity with mouse and rat ATP5H, respectively.
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Cross-reactivity
No cross-reactivity with other proteins
Reactive Species
PB9328 is reactive to ATP5H in Human, Monkey, Mouse, Rat
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Observed Molecular Weight
22 kDa
Calculated molecular weight
18491 MW
Background of ATP5PD
ATP5H is also known as ATPQ. Mitochondrial ATP synthase catalyzes ATP synthesis, utilizing an electrochemical gradient of protons across the inner membrane during oxidative phosphorylation. It is composed of two linked multi-subunit complexes: the soluble catalytic core, F1, and the membrane-spanning component, Fo, which comprises the proton channel. The F1 complex consists of 5 different subunits (alpha, beta, gamma, delta, and epsilon) assembled in a ratio of 3 alpha, 3 beta, and a single representative of the other 3. The Fo seems to have nine subunits (a, b, c, d, e, f, g, F6 and 8). This gene encodes the d subunit of the Fo complex. Alternatively spliced transcript variants encoding different isoforms have been identified for this gene. In addition, three pseudogenes are located on chromosomes 9, 12 and 15.
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
PB9328 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
Immunohistochemistry (Paraffin-embedded Section),2-5μg/ml, By Heat
Immunocytochemistry/Immunofluorescence, 5μg/ml
Positive Control
WB: human Hela whole cell, human HepG2 whole cell, human U-87MG whole cell, human A549 whole cell, monkey COS-7 whole cell, human SW620 whole cell, human Jurkat whole cell, human 293T whole cell, rat brain tissue, rat liver tissue, rat heart tissue, rat PC-12 whole cell, mouse brain tissue, mouse liver tissue, mouse heart tissue, mouse ANA-1 whole cell
IHC: human rectal cancer tissue
ICC/IF: T-47D cell
Validation Images & Assay Conditions
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Figure 1. Western blot analysis of ATP5H using anti-ATP5H antibody (PB9328).
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 Hela whole cell lysates,
Lane 2: human HepG2 whole cell lysates,
Lane 3: human U-87MG whole cell lysates,
Lane 4: human A549 whole cell lysates,
Lane 5: monkey COS-7 whole cell lysates,
Lane 6: human SW620 whole cell lysates,
Lane 7: human Jurkat whole cell lysates,
Lane 8: human 293T whole cell 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-ATP5H antigen affinity purified polyclonal antibody (Catalog # PB9328) 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 ATP5H at approximately 22 kDa. The expected band size for ATP5H is at 22 kDa.
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Figure 2. Western blot analysis of ATP5H using anti-ATP5H antibody (PB9328).
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: rat brain tissue lysates,
Lane 2: rat liver tissue lysates,
Lane 3: rat heart tissue lysates,
Lane 4: rat PC-12 whole cell lysates,
Lane 5: mouse brain tissue lysates,
Lane 6: mouse liver tissue lysates,
Lane 7: mouse heart tissue lysates,
Lane 8: mouse ANA-1 whole cell 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-ATP5H antigen affinity purified polyclonal antibody (Catalog # PB9328) 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 ATP5H at approximately 22 kDa. The expected band size for ATP5H is at 22 kDa.
Click image to see more details
Figure 3. IF analysis of ATP5H using anti-ATP5H antibody (PB9328).
ATP5H was detected in an immunocytochemical section of T-47D cells. 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 5 μg/mL rabbit anti-ATP5H Antibody (PB9328) 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.
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Figure 4. IHC analysis of ATP5H using anti-ATP5H antibody (PB9328).
ATP5H was detected in a paraffin-embedded section of human rectal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-ATP5H Antibody (PB9328) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Protein Target Info & Infographic
Gene/Protein Information For ATP5H (Source: Uniprot.org, NCBI)
Gene Name
ATP5H
Full Name
ATP synthase subunit d, mitochondrial
Weight
18491 MW
Superfamily
ATPase d subunit family
Alternative Names
ATP synthase subunit d, mitochondrial;ATPase subunit d;ATP5H;My032; ATP5PD APT5H, ATP5H, ATPQ ATP synthase peripheral stalk subunit d ATP synthase subunit d, mitochondrial|ATP synthase D chain, mitochondrial|ATP synthase, H+ transporting, mitochondrial F0 complex, subunit d|ATP synthase, H+ transporting, mitochondrial F1F0, subunit d|ATP synthase, H+ transporting, mitochondrial Fo complex subunit D|ATP synthase, H+ transporting, mitochondrial Fo complex, subunit d|ATPase subunit d|My032 protein
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on ATP5H, check out the ATP5H Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for ATP5H: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-ATP5H Antibody Picoband® (PB9328)
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Customer Q&As
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4 Customer Q&As for Anti-ATP5H Antibody Picoband®
Question
We are currently using anti-ATP5H antibody PB9328 for rat tissue, and we are happy with the IHC results. The species of reactivity given in the datasheet says human, mouse, rat. Is it true that the antibody can work on dog tissues as well?
Verified Customer
Verified customer
Asked: 2019-08-08
Answer
The anti-ATP5H antibody (PB9328) has not been tested for cross reactivity specifically with dog 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 dog you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2019-08-08
Question
Is a blocking peptide available for product anti-ATP5H antibody (PB9328)?
C. Collins
Verified customer
Asked: 2018-10-09
Answer
We do provide the blocking peptide for product anti-ATP5H antibody (PB9328). If you would like to place an order for it please contact [email protected] and make a special request.
Boster Scientific Support
Answered: 2018-10-09
Question
Can you help my question with product PB9328, anti-ATP5H antibody. I was wondering if it would be possible to conjugate this antibody with biotin. I would need it to be without BSA or sodium azide. I am planning on using a buffer exchange of sodium azide with PBS only. Would there be problems for me to conjugate the antibody and store it in -20 degrees in small aliquots?
R. Huang
Verified customer
Asked: 2016-05-18
Answer
We suggest not storing this antibody with PBS buffer only in -20 degrees. If you want to store it in -20 degrees it is best to add some cryoprotectant like glycerol. If you want carrier free PB9328 anti-ATP5H antibody, we can provide it to you in a special formula with trehalose and/or glycerol. These molecules will not interfere with conjugation chemistry and provide a good level of protection for the antibody from degradation. Please be sure to specify this in your purchase order.
Boster Scientific Support
Answered: 2016-05-18
Question
Here is the WB image, lot number and protocol we used for bone marrow {eco:0000313 using anti-ATP5H antibody PB9328. Please let me know if you require anything else.
S. Wu
Verified customer
Asked: 2013-03-04
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: 2013-03-04
