SKU PA1600-1
Size 100μg/vial
Reactivity Human, Mouse, Rat
Clonality Polyclonal
Host Rabbit
Ig Isotype N/A
Applications Flow Cytometry, IF, IHC-P, IHC-F, ICC, WB

Overview

Product Name Anti-HDAC3 Antibody
SKU/Catalog Number PA1600-1
Storage & Handling At -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time.Avoid repeated freezing and thawing.
Size 100μg/vial
Description Rabbit IgG polyclonal antibody for Histone deacetylase 3(HDAC3) detection. Tested with WB, IHC-P, IHC-F, ICC/IF, FCM in Human;Mouse;Rat.
Cite This Product Anti-HDAC3 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1600-1)
Host Rabbit
Contents/Buffer Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg Thimerosal, 0.05mg NaN3.
Form Lyophilized
Immunogen A synthetic peptide corresponding to a sequence at the C-terminus of human HDAC3(411-428aa NEFYDGDHDNDKESDVEI), identical to the related rat and mouse sequences.
Reactivity Human, Mouse, Rat

Assay Details

Assay Dilutions Overview

Concentration: Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Western blot, 0.1-0.5μg/ml, Human, Mouse, Rat
Immunohistochemistry(Paraffin-embedded Section), 0.5-1μg/ml, Human, Mouse, Rat, By Heat
Immunohistochemistry(Frozen Section), 0.5-1μg/ml, Human, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 2μg/ml, Human, Mouse, Rat
Flow Cytometry, 1-3μg/1x106 cells, Human

Boster's Secondary Antibodies And IHC, WB Kits

The following reagents are used to generate the images below.

Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit (SV0002-1) for IHC(P), IHC(F) and ICC.

Images And Assay Conditions

Figure 1. IHC analysis of HDAC3 using anti-HDAC3 antibody (PA1600-1).
HDAC3 was detected in paraffin-embedded section of human intestinal 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-HDAC3 Antibody (PA1600-1) 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.

Figure 2. IHC analysis of HDAC3 using anti-HDAC3 antibody (PA1600-1).
HDAC3 was detected in paraffin-embedded section of human lung 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-HDAC3 Antibody (PA1600-1) 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.

Figure 3. IHC analysis of HDAC3 using anti-HDAC3 antibody (PA1600-1).
HDAC3 was detected in paraffin-embedded section of Rat Intestine 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-HDAC3 Antibody (PA1600-1) 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.

Figure 4. IHC analysis of HDAC3 using anti-HDAC3 antibody (PA1600-1).
HDAC3 was detected in frozen section of Mouse Brain tissues. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-HDAC3 Antibody (PA1600-1) 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.

Figure 5. IHC analysis of HDAC3 using anti-HDAC3 antibody (PA1600-1).
HDAC3 was detected in immunocytochemical section of A549 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 1μg/ml rabbit anti-HDAC3 Antibody (PA1600-1) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

Figure 6. Western blot analysis of HDAC3 using anti-HDAC3 antibody (PA1600-1).
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 50ug of sample under reducing conditions.
Lane 1: Rat Stomach Tissue Lysate
Lane 2: Rat Testis Tissue Lysate
Lane 3: MCF-7 Cell Lysate
Lane 4: HELA Cell Lysate
Lane 5: JURKAT Cell Lysate
Lane 6: SKOV Cell Lysate
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-HDAC3 antigen affinity purified polyclonal antibody (Catalog # PA1600-1) 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 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system.

Figure 8. IF analysis of HDAC3 using anti- HDAC3 antibody (PA1600-1).
HDAC3 was detected in immunocytochemical section of Hela 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- HDAC3 Antibody (PA1600-1) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) 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.

Figure 7. IF analysis of HDAC3 using anti- HDAC3 antibody (PA1600-1).
HDAC3 was detected in immunocytochemical section of U20S 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- HDAC3 Antibody (PA1600-1) 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.

Figure 9. Flow Cytometry analysis of A431 cells using anti-HDAC3 antibody (PA1600-1).
Overlay histogram showing A431 cells stained with PA1600-1 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti- HDAC3 Antibody (PA1600-1,1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Figure 10. Flow Cytometry analysis of U937cells using anti-HDAC3 antibody (PA1600-1).
Overlay histogram showing U937 cells stained with PA1600-1 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti- HDAC3 Antibody (PA1600-1, 1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Target Info

Protein Target Info (Source: Uniprot.org)

Uniprot Id O15379
Gene Name HDAC3
Protein Name Histone deacetylase 3
Tissue Specificity Widely expressed.
Alternative Names Histone deacetylase 3;HD3;3.5.1.98;RPD3-2;SMAP45;HDAC3;
Subcellular Localization Nucleus . Cytoplasm . Cytoplasm, cytosol . Colocalizes with XBP1 and AKT1 in the cytoplasm (PubMed:25190803). Predominantly expressed in the nucleus in the presence of CCAR2. .
Molecular Weight 48848 MW

*if product is indicated to react with multiple species, protein info is based on the human gene.

Ontology

Protein Function Responsible for the deacetylation of lysine residues on the N-terminal part of the core histones (H2A, H2B, H3 and H4), and some other non-histone substrates. Histone deacetylation gives a tag for epigenetic repression and plays an important role in transcriptional regulation, cell cycle progression and developmental events. Histone deacetylases act via the formation of large multiprotein complexes. Participates in the BCL6 transcriptional repressor activity by deacetylating the H3 'Lys- 27' (H3K27) on enhancer elements, antagonizing EP300 acetyltransferase activity and repressing proximal gene expression. Probably participates in the regulation of transcription through its binding to the zinc-finger transcription factor YY1; increases YY1 repression activity. Required to repress transcription of the POU1F1 transcription factor. Acts as a molecular chaperone for shuttling phosphorylated NR2C1 to PML bodies for sumoylation (PubMed:21444723, PubMed:23911289). Contributes, together with XBP1 isoform 1, to the activation of NFE2L2-mediated HMOX1 transcription factor gene expression in a PI(3)K/mTORC2/Akt-dependent signaling pathway leading to endothelial cell (EC) survival under disturbed flow/oxidative stress (PubMed:25190803). .
Research Areas Human, Mouse, Rat

*You can search these to find other products in these research areas.
Background HDAC3(HISTONE DEACETYLASE 3) is a member of the histone deacetylase/acuc/apha family of proteins that is an enzyme that in humans is encoded by the HDAC3 gene. The HDAC3 gene is mapped to 5q31.3. HDAC3 has histone deacetylase activity and represses transcription when tethered to a promoter. It may participate in the regulation of transcription through its binding with the zinc-finger transcription factor YY1. The protein can also down-regulate p53 function and thus modulate cell growth and apoptosis. And this gene is regarded as a potential tumor suppressor gene. HDAC3 has an open reading frame of 428 amino acids and shares 53% amino acid identity with HDAC1 and 52% with HDAC2. The catalytic domain of HDAC4 interacts with HDAC3 via the transcriptional corepressor NCOR2. All experimental conditions leading to the suppression of HDAC4 binding to NCOR2 and to HDAC3 resulted in loss of enzymatic activity associated with HDAC4. HDAC3 recruitment to the genome displays a circadian rhythm in mouse liver.

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Polyclonal antibody for HDAC3 detection. Host: Rabbit.Size: 100μg/vial. Tested applications: WB, IHC-P, IHC-F, ICC/IF, FCM. Reactive species: Human. HDAC3 information: Molecular Weight: 48848 MW; Subcellular Localization: Nucleus . Cytoplasm . Cytoplasm, cytosol . Colocalizes with XBP1 and AKT1 in the cytoplasm (PubMed:25190803). Predominantly expressed in the nucleus in the presence of CCAR2; Tissue Specificity: Widely expressed.
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PA1600-1
Buy one primary antibody get one 0.5ml HRP or Biotin secondary antibody for free.
*Sample sizes are prepared on demand and will take extra lead time. (cannot be conjugated)
$280.00

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Publications

Interleukin-18 down-regulates multidrug resistance-associated protein 2 expression through farnesoid x receptor associated with nuclear factor kappa B and %u2026
Mitochondrial BNIP3 upregulation precedes endonuclease G translocation in hippocampal neuronal death following oxygen-glucose deprivation

Customer Q&As

  • Q: Do you offer BSA-free antibodies? Keyword: Bovine serum albumin, carrier protein, conjugation
    A: Yes, please contact us at support@bosterbio.com for more information about BSA-free antibodies and availability. The new BSA-free formula uses trehalose as a replacement to BSA. We have tested many alternative chemicals and found that trehalose protects the antibodies the best.
  • Q: Is your western blot protocol provided from the website applicable for all your antibodies? Keyword: applications, WB
    A: The protocol is applicable for all our antibodies in WB, the NC Membrane(0.45μm or 0.22μm) and transfer time(70 mins or 50 mins) depends on the protein molecular weight, details can be found in included protocol.
  • Q: Can I conjugate markers to this antibody? Can I link custom conjugates to this antibody? Keyword: conjugation
    A: The antibody is stored with BSA and cannot be conjugated with markers. Carrier free antibodies are available upon request. Please contact support@bosterbio.com
  • Q: What should I use for negative control?
    A: Please contact us for negative control suggestions. You can also check expression databases such as genecards, uniprot etc. Due to logistic reasons, we do not sell serum or lysates that we use internally for positive or negative control.
  • Q: Where can I find troubleshooting information? What should I do if I have unexpected bands, high background, no signal, weak signal
    A: You can find Boster's troubleshoot guides under tech support tab. Please contact us for further assistance on troubleshooting your experiment.
  • Q: What is the immunogen sequence of this antibody? Is this antibody polyclonal or monoclonal?
    A: You can find the immunogen sequence under "
  • Q: What is the expected band size? Why is it different than the observed band size?
    A: The expected band size is predicted on the size of the protein. The actual band size may be affected by a few other factors including but not limited to:<br>1. Post-translational modification:phosphorylation, methylation, glycosylation etc. These modifications prevent SDS molecules from binding to the target protein and thus make the band size appear larger than expected<br>2. Post-translational cleavage: this can cause smaller bands and or multiple bands <br><br>3. Alternative splicing: the same gene can have alternative splicing patterns generating different size proteins, all with reactivities to the antibody. <br><br>4. Amino Acid R chain charge: SDS binds to positive charges. The different size and charge of the Amino Acid side chains can affect the amount of SDS binding and thus affect the observed band size.<br>5. Multimers: Multimers are usually broken up in reducing conditions. However if the interactions between the multimers are strong, the band may appear higher., <br>
  • Q: What is the suggested dilution ratio for Western Blot (WB), Immunohistochemistry (IHC) and or ELISA standards? What is the optimal pH for the sample?
    A: Check the datasheet for the product for details on dilution ratios for different experiments. You can find the datasheet button on the right side of the product page.
  • Q: What is the protocol you used for your Western blotting (WB) and Immunohistochemistry (IHC)?
    A: Check our protocols under the tech support tab.
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