Recombinant Anti-KAT2A / GCN5 antibody [EPR21146] - ChIP Grade (ab217876)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR21146] to KAT2A / GCN5 - ChIP Grade
- Suitable for: ChIP, WB, IP
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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Product name
Anti-KAT2A / GCN5 antibody [EPR21146] - ChIP Grade
See all KAT2A / GCN5 primary antibodies -
Description
Rabbit monoclonal [EPR21146] to KAT2A / GCN5 - ChIP Grade -
Host species
Rabbit -
Tested applications
Suitable for: ChIP, WB, IPmore details -
Species reactivity
Reacts with: Mouse, Human
Predicted to work with: Rat -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HAP1, HEK-293T, MCF7 and HeLa whole cell lysate; human fetal brain tissue lysate; His-tagged human KAT2A / GCN5 recombinant protein (aa86-336); Wild-type U-2 OS and NIH/3T3 cell lysates. IP: HeLa whole cell lysate.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR21146 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab217876 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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ChIP |
Use at an assay dependent concentration.
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WB |
1/1000. Detects a band of approximately 94 kDa (predicted molecular weight: 94 kDa).
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IP |
1/30.
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Notes |
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ChIP
Use at an assay dependent concentration. |
WB
1/1000. Detects a band of approximately 94 kDa (predicted molecular weight: 94 kDa). |
IP
1/30. |
Target
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Function
Functions as a histone acetyltransferase (HAT) to promote transcriptional activation. Acetylation of histones gives a specific tag for epigenetic transcription activation. Has significant histone acetyltransferase activity with core histones, but not with nucleosome core particles. In case of HIV-1 infection, it is recruited by the viral protein Tat. Regulates Tat's transactivating activity and may help inducing chromatin remodeling of proviral genes. Component of the ATAC complex, a complex with histone acetyltransferase activity on histones H3 and H4. -
Tissue specificity
Expressed in all tissues tested, with most abundant expression in ovary. -
Sequence similarities
Belongs to the GCN5 family.
Contains 1 bromo domain.
Contains 1 N-acetyltransferase domain. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 2648 Human
- Entrez Gene: 14534 Mouse
- Entrez Gene: 303539 Rat
- Omim: 602301 Human
- SwissProt: Q92830 Human
- SwissProt: Q9JHD2 Mouse
- Unigene: 463045 Human
- Unigene: 218837 Mouse
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Alternative names
- 1110051E14Rik antibody
- AW212720 antibody
- EC 2.3.1.48 antibody
see all
Images
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All lanes : Anti-KAT2A / GCN5 antibody [EPR21146] - ChIP Grade (ab217876) at 1/1000 dilution
Lane 1 : Wild-type U-2 OS cell lysate
Lane 2 : GCN5 knockout U-2 OS cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 94 kDa
Observed band size: 94 kDaWestern blot: Anti-KAT2A / GCN5 antibody [EPR21146] - ChIP Grade staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab217876 was shown to bind specifically to KAT2A / GCN5. A band was observed at 94 kDa in wild-type U-2 OS cell lysates with no signal observed at this size in kat2a knockout cell line. To generate this image, wild-type and kat2a knockout U-2 OS cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
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All lanes : Anti-KAT2A / GCN5 antibody [EPR21146] - ChIP Grade (ab217876) at 1/1000 dilution
Lane 1 : Wild-type U-2 OS cell lysate
Lane 2 : GCN5 knockout U-2 OS cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : NIH/3T3 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 94 kDa
Observed band size: 95 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-KAT2A / GCN5 antibody [EPR21146] - ChIP Grade staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab217876 was shown to bind specifically to KAT2A / GCN5. A band was observed at 95 kDa in wild-type U-2 OS cell lysates with no signal observed at this size in kat2a knockout cell line. To generate this image, wild-type and kat2a knockout U-2 OS cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
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All lanes : Anti-KAT2A / GCN5 antibody [EPR21146] - ChIP Grade (ab217876) at 1/1000 dilution
Lane 1 : His-tagged human KAT2A / GCN5 recombinant protein (aa86-336), 10 ng
Lane 2 : His-tagged human KAT2B recombinant protein (aa74-326), 10 ng
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 94 kDa
Observed band size: 27 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 1 second
The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument.
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Chromatin was prepared from A549 treated with thapsigargin (1µM 12 hours) cells according to the Abcam Dual X-ChIP protocol*. Cells were fixed with EGS for 30 minutes, then formaldehyde for 10 minutes.
The ChIP was performed with 25 µg of chromatin, 5 µg of ab217876 (red), and 20 µl of Protein A/G sepharose beads. 5 µg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
Primers and probes are located in the first kb of the transcribed region.
*http://www.abcam.com/resources?keywords=X%20ChIP%20protocol -
KAT2A / GCN5 was immunoprecipitated from 0.35mg of HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab217876 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab217876 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.
Lane 1: HeLa whole cell lysate 10ug (Input).
Lane 2: ab217876 IP in HeLa whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab217876 in HeLa whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 seconds. -
All lanes : Anti-KAT2A / GCN5 antibody [EPR21146] - ChIP Grade (ab217876) at 1/1000 dilution
Lane 1 : Wild type HAP1 whole cell lysate
Lane 2 : KAT2A / GCN5-knockout HAP1 whole cell lysate whole cell lysate
Lane 3 : HEK-293T (uman epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
Lane 4 : MCF7 (human breast adenocarcinoma cell line) whole cell lysate
Lysates/proteins at 10 µg per lane.
Predicted band size: 94 kDa
Observed band size: 94 kDa
Exposure time: 3 minutesab217876 was shown to specifically react with KAT2A / GCN5 in wild-type HAP1 cells as signal was lost in KAT2A / GCN5 knockout cells. Wild-type and KAT2A / GCN5 knockout samples were subjected to SDS-PAGE. Ab217876 and ab181602 (Human anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/200000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™ MP instrumentusing the ECL technique.
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All lanes : Anti-KAT2A / GCN5 antibody [EPR21146] - ChIP Grade (ab217876) at 1/1000 dilution
Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : Human fetal brain tissue lysate
Lysates/proteins at 10 µg per lane.
Secondary
Lane 1 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Lane 2 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Predicted band size: 94 kDa
Observed band size: 94 kDaBlocking/Dilution buffer: 5% NFDM/TBST
Exposure time: Lane 1: 15 seconds; Lane 2: 3 minutes
The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (6)
ab217876 has been referenced in 6 publications.
- He B et al. Long noncoding RNA LINC00930 promotes PFKFB3-mediated tumor glycolysis and cell proliferation in nasopharyngeal carcinoma. J Exp Clin Cancer Res 41:77 (2022). PubMed: 35209949
- Tian X et al. The miR-5694/AF9/Snail Axis Provides Metastatic Advantages and a Therapeutic Target in Basal-like Breast Cancer. Mol Ther 29:1239-1257 (2021). PubMed: 33221433
- Guo Y et al. Oncogenic Chromatin Modifier KAT2A Activates MCT1 to Drive the Glycolytic Process and Tumor Progression in Renal Cell Carcinoma. Front Cell Dev Biol 9:690796 (2021). PubMed: 34268311
- Lei I et al. Acetyl-CoA production by specific metabolites promotes cardiac repair after myocardial infarction via histone acetylation. Elife 10:N/A (2021). PubMed: 34939931
- Liao W et al. Upregulation of GPNCA is associated with poor prognosis through enhancement of tumor growth via regulating GSK3B. Sci Rep 10:2044 (2020). PubMed: 32029792
- Cheng Z et al. K-Ras-ERK1/2 accelerates lung cancer cell development via mediating H3K18ac through the MDM2-GCN5-SIRT7 axis. Pharm Biol 57:701-709 (2019). PubMed: 31613681