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Product Info Summary
| SKU: | PB9628 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | Flow Cytometry, IHC, WB |
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Product info
Product Name
Anti-HDAC6 Antibody Picoband®
SKU/Catalog Number
PB9628
PB0672 is an alternative SKU for this antibody, used in previous lots.
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-HDAC6 Antibody Picoband® catalog # PB9628. Tested in Flow Cytometry, IHC, WB applications. This antibody reacts with Human, 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-HDAC6 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9628)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
A synthetic peptide corresponding to a sequence at the N-terminus of human HDAC6, different from the related mouse sequence by one amino acid.
Cross-reactivity
No cross-reactivity with other proteins
Reactive Species
PB9628 is reactive to HDAC6 in Human, Mouse, Rat
Observed Molecular Weight
160 kDa
Calculated molecular weight
131.4 kDa
Background of HDAC6
HDAC6, also called KIAA0901, is a member belongs to class II of the histone deacetylase/acuc/apha family of proteins that is an enzyme that in humans is encoded by the HDAC6 gene. The HDAC6 gene is mapped to chromosome Xp11.23. HDAC6 contains an internal duplication of two catalytic domains which appear to function independently of each other. The protein possesses histone deacetylase activity and represses transcription. HDAC6 functions as a tubulin deacetylase. And it is localized exclusively in the cytoplasm, where it associates with microtubules and localizes with the microtubule motor complex. HDAC6 could bind both polyubiquitinated misfolded proteins and dynein motors, thereby recruiting misfolded protein cargo to dynein motors for transport to aggresomes. Furthermore, expression of HDAC6 was sufficient to rescue degeneration associated with UPS dysfunction in vivo in an autophagy-dependent manner. HDAC6 is a central component of the stress response that regulates SG formation and potentially contributes to control of RNA metabolism and translation.
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
PB9628 is guaranteed for Flow Cytometry, IHC, 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), 2-5 μg/ml, Human
Flow Cytometry(Fixed), 1-3 μg/1x106 cells, Human
Positive Control
WB: human K562 whole cell lysates, human HepG2 whole cell lysates, human SH-SY5Y whole cell lysates, rat brain tissue lysates, mouse brain tissue lysates
IHC: human breast cancer tissue
FCM: K562 cell
Validation Images & Assay Conditions
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Western blot analysis of HDAC6 using anti-HDAC6 antibody (PB9628).
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 K562 whole cell lysates,
Lane 2: human HepG2 whole cell lysates,
Lane 3: human SH-SY5Y whole cell lysates,
Lane 4: rat brain tissue lysates,
Lane 5: mouse brain 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-HDAC6 antigen affinity purified polyclonal antibody (Catalog # PB9628) 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 HDAC6 at approximately 160 kDa. The expected band size for HDAC6 is at 131 kDa.
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IHC analysis of HDAC6 using anti-HDAC6 antibody (PB9628).
HDAC6 was detected in a paraffin-embedded section of human breast 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-HDAC6 Antibody (PB9628) 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.
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IHC analysis of HDAC6 using anti-HDAC6 antibody (PB9628).
HDAC6 was detected in a paraffin-embedded section of human breast 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-HDAC6 Antibody (PB9628) 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.
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Flow Cytometry analysis of K562 cells using anti-HDAC6 antibody (PB9628).
Overlay histogram showing K562 cells stained with PB9628 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-HDAC6 Antibody (PB9628, 1 μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Specific Publications For Anti-HDAC6 Antibody Picoband® (PB9628)
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4 Customer Q&As for Anti-HDAC6 Antibody Picoband®
Question
We have seen staining in rat brain. Do you have any suggestions? Is anti-HDAC6 antibody supposed to stain brain positively?
Verified Customer
Verified customer
Asked: 2017-10-02
Answer
From what I have seen in literature brain does express HDAC6. From what I have seen in Uniprot.org, HDAC6 is expressed in adenohypophysis, brain, ovary placenta, cervix carcinoma erythroleukemia, liver, among other tissues. Regarding which tissues have HDAC6 expression, here are a few articles citing expression in various tissues:
Brain, Pubmed ID: 10048485, 15772651
Cervix carcinoma, and Erythroleukemia, Pubmed ID: 23186163
Liver, Pubmed ID: 24275569
Ovary, and Placenta, Pubmed ID: 15489334
Boster Scientific Support
Answered: 2017-10-02
Question
I am interested in using your anti-HDAC6 antibody for polyubiquitinated misfolded protein transport studies. Has this antibody been tested with western blotting on rat skeletal muscle tissue? We would like to see some validation images before ordering.
Verified Customer
Verified customer
Asked: 2017-06-16
Answer
We appreciate your inquiry. This PB9628 anti-HDAC6 antibody is tested on rat skeletal muscle tissue, tissue lysate, colo320 whole cell lysate. It is guaranteed to work for WB in human, rat. Our Boster guarantee will cover your intended experiment even if the sample type has not been be directly tested.
Boster Scientific Support
Answered: 2017-06-16
Question
Our team were satisfied with the WB result of your anti-HDAC6 antibody. However we have been able to see positive staining in ovary placenta nucleus. cytoplasm. perikaryon using this antibody. Is that expected? Could you tell me where is HDAC6 supposed to be expressed?
W. Singh
Verified customer
Asked: 2015-09-16
Answer
According to literature, ovary placenta does express HDAC6. Generally HDAC6 expresses in nucleus. cytoplasm. perikaryon. Regarding which tissues have HDAC6 expression, here are a few articles citing expression in various tissues:
Brain, Pubmed ID: 10048485, 15772651
Cervix carcinoma, and Erythroleukemia, Pubmed ID: 23186163
Liver, Pubmed ID: 24275569
Ovary, and Placenta, Pubmed ID: 15489334
Boster Scientific Support
Answered: 2015-09-16
Question
We are currently using anti-HDAC6 antibody PB9628 for rat tissue, and we are happy with the WB results. The species of reactivity given in the datasheet says human, rat. Is it likely that the antibody can work on bovine tissues as well?
M. Kulkarni
Verified customer
Asked: 2013-07-09
Answer
The anti-HDAC6 antibody (PB9628) has not been tested for cross reactivity specifically with bovine tissues, though 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 bovine you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2013-07-09
