Recombinant Anti-KAT2A / GCN5 antibody [EPR21146] - BSA and Azide free (ab231075)
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 - BSA and Azide free
- Suitable for: ChIP, WB, IP
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
-
Product name
Anti-KAT2A / GCN5 antibody [EPR21146] - BSA and Azide free
See all KAT2A / GCN5 primary antibodies -
Description
Rabbit monoclonal [EPR21146] to KAT2A / GCN5 - BSA and Azide free -
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.
-
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.
-
General notes
ab231075 is the carrier-free version of ab217876.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
-
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR21146 -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Conjugation kits
-
Isotype control
-
Recombinant Protein
-
Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab231075 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 |
---|---|---|
ChIP |
Use at an assay dependent concentration.
|
|
WB |
Use at an assay dependent concentration. Detects a band of approximately 94 kDa (predicted molecular weight: 94 kDa).
|
|
IP |
Use at an assay dependent concentration.
|
Notes |
---|
ChIP
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Detects a band of approximately 94 kDa (predicted molecular weight: 94 kDa). |
IP
Use at an assay dependent concentration. |
Target
-
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
-
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
-
Alternative names
- 1110051E14Rik antibody
- AW212720 antibody
- EC 2.3.1.48 antibody
see all
Images
-
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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab217876).
-
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.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 94 kDaBlocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 3 minutes
ab217876 was shown to specifically react with KAT2A in wild type HAP1 cells. No band was observed when KAT2A knockout samples were used. Wild-type and KAT2A knockout samples were subjected to SDS-PAGE. ab217876 and ab181602 (Human anti-GAPDH loading control) were incubated overnight at 4°C at a 1/1000 dilution and 1/5000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab217876).
-
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%20protocolThis data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab217876).
-
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.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab217876).
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
-
Datasheet download
Certificate of Compliance
References (2)
ab231075 has been referenced in 2 publications.
- Liu Y et al. Cyclic Mechanical Strain Regulates Osteoblastic Differentiation of Mesenchymal Stem Cells on TiO2 Nanotubes Through GCN5 and Wnt/β-Catenin. Front Bioeng Biotechnol 9:735949 (2021). PubMed: 34869255
- Bao X et al. Glutarylation of Histone H4 Lysine 91 Regulates Chromatin Dynamics. Mol Cell 76:660-675.e9 (2019). PubMed: 31542297