Recombinant Anti-KAT3B / p300 antibody [EPR23495-268] - ChIP Grade (ab275378)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23495-268] to KAT3B / p300 - ChIP Grade
- Suitable for: WB, IP, ChIP, Flow Cyt (Intra), ICC/IF
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-KAT3B / p300 antibody [EPR23495-268] - ChIP Grade
See all KAT3B / p300 primary antibodies -
Description
Rabbit monoclonal [EPR23495-268] to KAT3B / p300 - ChIP Grade -
Host species
Rabbit -
Specificity
Anti-KAT3B / p300 antibody [EPR23495-268] reacts weakly with CREBBP.
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Tested applications
Suitable for: WB, IP, ChIP, Flow Cyt (Intra), ICC/IFmore details
Unsuitable for: IHC-P -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: K-562, RAW 264.7 and PC-12 whole cell lysates; His-tagged human KAT3B/p300 recombinant protein. ICC/IF: HeLa and NIH/3T3 cells. Flow Cyt (intra): NIH/3T3 cells. IP: K-562 whole cell lysate. ChIP: Chromatin prepared from K-562 cells.
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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
Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR23495-268 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab275378 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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WB | (1) |
1/1000. Detects a band of approximately 300 kDa (predicted molecular weight: 264 kDa).
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IP |
1/30.
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ChIP |
Use a concentration of 5 µg/ml.
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Flow Cyt (Intra) |
1/500.
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ICC/IF |
1/100.
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Notes |
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WB
1/1000. Detects a band of approximately 300 kDa (predicted molecular weight: 264 kDa). |
IP
1/30. |
ChIP
Use a concentration of 5 µg/ml. |
Flow Cyt (Intra)
1/500. |
ICC/IF
1/100. |
Target
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Function
Functions as histone acetyltransferase and regulates transcription via chromatin remodeling. Acetylates all four core histones in nucleosomes. Histone acetylation gives an epigenetic tag for transcriptional activation. Mediates cAMP-gene regulation by binding specifically to phosphorylated CREB protein. Mediates acetylation of histone H3 at 'Lys-122' (H3K122ac), a modification that localizes at the surface of the histone octamer and stimulates transcription, possibly by promoting nucleosome instability. Mediates acetylation of histone H3 at 'Lys-27' (H3K27ac). Also functions as acetyltransferase for nonhistone targets. Acetylates 'Lys-131' of ALX1 and acts as its coactivator. Acetylates SIRT2 and is proposed to indirectly increase the transcriptional activity of TP53 through acetylation and subsequent attenuation of SIRT2 deacetylase function. Acetylates HDAC1 leading to its inactivation and modulation of transcription. Acts as a TFAP2A-mediated transcriptional coactivator in presence of CITED2. Plays a role as a coactivator of NEUROD1-dependent transcription of the secretin and p21 genes and controls terminal differentiation of cells in the intestinal epithelium. Promotes cardiac myocyte enlargement. Can also mediate transcriptional repression. Binds to and may be involved in the transforming capacity of the adenovirus E1A protein. 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. Acetylates FOXO1 and enhances its transcriptional activity. Acetylates BCL6 wich disrupts its ability to recruit histone deacetylases and hinders its transcriptional repressor activity. Participates in CLOCK or NPAS2-regulated rhythmic gene transcription; exhibits a circadian association with CLOCK or NPAS2, correlating with increase in PER1/2 mRNA and histone H3 acetylation on the PER1/2 promoter. Acetylates MTA1 at 'Lys-626' which is essential for its transcriptional coactivator activity (PubMed:10733570, PubMed:11430825, PubMed:11701890, PubMed:12402037, PubMed:12586840, PubMed:12929931, PubMed:14645221, PubMed:15186775, PubMed:15890677, PubMed:16617102, PubMed:16762839, PubMed:18722353, PubMed:18995842, PubMed:23415232, PubMed:23911289, PubMed:23934153, PubMed:8945521). Acetylates XBP1 isoform 2; acetylation increases protein stability of XBP1 isoform 2 and enhances its transcriptional activity (PubMed:20955178). Acetylates PCNA; acetylation promotes removal of chromatin-bound PCNA and its degradation during nucleotide excision repair (NER) (PubMed:24939902). Acetylates MEF2D. -
Involvement in disease
Defects in EP300 may play a role in epithelial cancer.
Chromosomal aberrations involving EP300 may be a cause of acute myeloid leukemias. Translocation t(8;22)(p11;q13) with KAT6A.
Rubinstein-Taybi syndrome 2 -
Sequence similarities
Contains 1 bromo domain.
Contains 1 CBP/p300-type HAT (histone acetyltransferase) domain.
Contains 1 KIX domain.
Contains 2 TAZ-type zinc fingers.
Contains 1 ZZ-type zinc finger. -
Domain
The CRD1 domain (cell cycle regulatory domain 1) mediates transcriptional repression of a subset of p300 responsive genes; it can be de-repressed by CDKN1A/p21WAF1 at least at some promoters. It conatins sumoylation and acetylation sites and the same lysine residues may be targeted for the respective modifications. It is proposed that deacetylation by SIRT1 allows sumoylation leading to suppressed activity. -
Post-translational
modificationsAcetylated on Lys at up to 17 positions by intermolecular autocatalysis. Deacetylated in the transcriptional repression domain (CRD1) by SIRT1, preferentially at Lys-1020. Deacetylated by SIRT2, preferentially at Lys-418, Lys-423, Lys-1542, Lys-1546, Lys-1549, Lys-1699, Lys-1704 and Lys-1707.
Citrullinated at Arg-2142 by PADI4, which impairs methylation by CARM1 and promotes interaction with NCOA2/GRIP1.
Methylated at Arg-580 and Arg-604 in the KIX domain by CARM1, which blocks association with CREB, inhibits CREB signaling and activates apoptotic response. Also methylated at Arg-2142 by CARM1, which impairs interaction with NCOA2/GRIP1.
Sumoylated; sumoylation in the transcriptional repression domain (CRD1) mediates transcriptional repression. Desumoylated by SENP3 through the removal of SUMO2 and SUMO3.
Probable target of ubiquitination by FBXO3, leading to rapid proteasome-dependent degradation.
Phosphorylated by HIPK2 in a RUNX1-dependent manner. This phosphorylation that activates EP300 happens when RUNX1 is associated with DNA and CBFB. Phosphorylated by ROCK2 and this enhances its activity. Phosphorylation at Ser-89 by AMPK reduces interaction with nuclear receptors, such as PPARG. -
Cellular localization
Cytoplasm. Nucleus. In the presence of ALX1 relocalizes from the cytoplasm to the nucleus. Colocalizes with ROCK2 in the nucleus. - Information by UniProt
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Database links
- Entrez Gene: 2033 Human
- Entrez Gene: 328572 Mouse
- Entrez Gene: 170915 Rat
- Omim: 602700 Human
- SwissProt: Q09472 Human
- SwissProt: B2RWS6 Mouse
- Unigene: 517517 Human
- Unigene: 655211 Human
see all -
Alternative names
- E1A associated protein p300 antibody
- E1A binding protein p300 antibody
- E1A-associated protein p300 antibody
see all
Images
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KAT3B/p300 was immunoprecipitated from 0.35 mg K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate with ab275378 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab275378 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate 10 ug
Lane 2: ab275378 IP in K-562 whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab275378 in K-562 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 76 seconds.
Lysate was freshly prepared and used in IP immediately to mininize protein degradation.
Incubation time was 2 hours.
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Chromatin was prepared from K-562 cells according to the Abcam Dual-X-ChIP protocol*. Cells were fixed with 1.5 mM EGS for 30mins and then formaldehyde for 10min.
The ChIP was performed with 25 µg of chromatin, 5 µg of ab275378 (red), or 5 µg of rabbit normal IgG ab172730 (gray) and 25 µl of Protein A/G sepharose beads. The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).*https://www.abcam.com/resources?keywords=X%20ChIP%20protocol
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 cells labelling KAT3B/p300 with ab275378 at 1/100 (4.77 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing nuclear and weakly cytoplasmic staining in NIH/3T3 cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized NIH/3T3 (Mouse embryonic fibroblast) cells labelling KAT3B/p300 with ab275378 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).
A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
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All lanes : Anti-KAT3B / p300 antibody [EPR23495-268] - ChIP Grade (ab275378) at 1/1000 dilution
Lane 1 : His-tagged human KAT3B/p300 recombinant protein 20 ng
Lane 2 : His-tagged human CREBBP recombinant protein, 20 ng
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution
Predicted band size: 264 kDa
Observed band size: 46.3,88.7 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Both recombinant proteins were expressed from a E.coil expression system. Sample loaded onto lane 2 was E.coil extract containing CREBBP recombinant protein.
This product has weak cross reaction with CREBBP protein.
Exposure time: 5.5 seconds.
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa cells labelling KAT3B/p300 with ab275378 at 1/100 (4.77 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing nuclear and weakly cytoplasmic staining in HeLa cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
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All lanes : Anti-KAT3B / p300 antibody [EPR23495-268] - ChIP Grade (ab275378) at 1/1000 dilution
Lane 1 : K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate
Lane 2 : RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 3 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution
Predicted band size: 264 kDa
Observed band size: 300 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The molecular weight observed is consistent with what has been described in the literature (PMID: 8995708).
Lysates were made freshly and used in WB immediately to minimize protein degradation.
Exposure times: Lane 1: 125 seconds;Lanes 2-3: 3 minutes.
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 (8)
ab275378 has been referenced in 8 publications.
- Liu Y et al. The CTCF/LncRNA-PACERR complex recruits E1A binding protein p300 to induce pro-tumour macrophages in pancreatic ductal adenocarcinoma via directly regulating PTGS2 expression. Clin Transl Med 12:e654 (2022). PubMed: 35184402
- Sherman EJ et al. Identification of cell type specific ACE2 modifiers by CRISPR screening. PLoS Pathog 18:e1010377 (2022). PubMed: 35231079
- Wang X et al. Combination regimen of granulocyte colony-stimulating factor and recombinant human thrombopoietin improves the curative effect on elderly patients with leukemia through inducing pyroptosis and ferroptosis of leukemia cells. Cancer Gene Ther 29:1742-1750 (2022). PubMed: 35768562
- Leng S et al. Ion channel Piezo1 activation promotes aerobic glycolysis in macrophages. Front Immunol 13:976482 (2022). PubMed: 36119083
- Shen W et al. Transcription Factor p300 Regulated miR-451b Weakens the Cigarette Smoke Extract-Induced Cellular Stress by Targeting RhoA/ROCK2 Signaling. Oxid Med Cell Longev 2022:7056283 (2022). PubMed: 36275894
- Lin J et al. Overexpression of Tfap2a in Mouse Oocytes Impaired Spindle and Chromosome Organization. Int J Mol Sci 23:N/A (2022). PubMed: 36430853
- Wang Z et al. CREB stimulates GPX4 transcription to inhibit ferroptosis in lung adenocarcinoma. Oncol Rep 45:N/A (2021). PubMed: 33846793
- Fan Z et al. An E2F5-TFDP1-BRG1 Complex Mediates Transcriptional Activation of MYCN in Hepatocytes. Front Cell Dev Biol 9:742319 (2021). PubMed: 34746136