Recombinant Anti-KAT3B / p300 antibody [EPR23753-55] (ab259330)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23753-55] to KAT3B / p300
- Suitable for: WB, ICC/IF, Flow Cyt (Intra), ChIC/CUT&RUN-seq
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
-
Product name
Anti-KAT3B / p300 antibody [EPR23753-55]
See all KAT3B / p300 primary antibodies -
Description
Rabbit monoclonal [EPR23753-55] to KAT3B / p300 -
Host species
Rabbit -
Tested applications
Suitable for: WB, ICC/IF, Flow Cyt (Intra), ChIC/CUT&RUN-seqmore details
Unsuitable for: ChIP,IHC-P or IP -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
-
Positive control
- WB: K562, His-tagged human EP300 recombinant protein, HeLa, K562, RAW264.7 and PC-12 whole cell lysate. ICC/IF: HeLa and NIH/3T3 cells. Flow Cyt: HeLa and NIH/3T3 cells. ChIC/CUT&RUN seq: HeLa cell.
-
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
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
-
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR23753-55 -
Isotype
IgG -
Research areas
Associated products
-
Compatible Secondaries
-
Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab259330 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 |
---|---|---|
WB |
1/1000. Predicted molecular weight: 264 kDa.
|
|
ICC/IF |
1/500.
|
|
Flow Cyt (Intra) |
1/500.
|
|
ChIC/CUT&RUN-seq |
Use at an assay dependent concentration.
|
Notes |
---|
WB
1/1000. Predicted molecular weight: 264 kDa. |
ICC/IF
1/500. |
Flow Cyt (Intra)
1/500. |
ChIC/CUT&RUN-seq
Use at an assay dependent concentration. |
Target
-
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
-
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
-
ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/µL, 2.5 x 10^5 HeLa (Human cervix adenocarcinoma epithelial cell line) cells and 5 µg of ab259330 [EPR23753-55]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.
Additional screenshots of mapped reads can be downloaded here. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods. -
All lanes : Anti-KAT3B / p300 antibody [EPR23753-55] (ab259330) at 1/1000 dilution
Lane 1 : K562 (human chronic myelogenous leukemia lymphoblast), whole cell lysate
Lane 2 : HCT15 (human colon epithelial cell), 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
Fresh lysates were used in this WB.
The ~100KDa signal is a non-target band.
Negative control: HCT15 (PMID:14732695)
Exposure time: 3 minutes
-
All lanes : Anti-KAT3B / p300 antibody [EPR23753-55] (ab259330) at 1/1000 dilution
Lane 1 : His-tagged human EP300 recombinant protein
Lane 2 : His-tagged human CREBBP recombinant protein
Lysates/proteins at 0.01 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution
Predicted band size: 264 kDaBlocking and diluting buffer and concentration: 5% NFDM/TBST
This antibody has no cross-reaction with human CREBBP.
Both recombinant proteins were made in-house.
The sample loaded onto lane 1 was purified EP300 recombinant protein expressed from an E.coli expression system.
The sample loaded onto lane 2 is E.coli extracts containing recombinant CREBBP protein.
Exposure time: 15 seconds
-
Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized HeLa cells labelling KAT3B / p300 with ab259330 at 1/500 (0.926 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green). Confocal image showing mainly nuclear staining in HeLa cells and no staining in HCT-15 cells. Negative control: HCT-15 (PMID: 14732695) is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/ml dilution.
-
All lanes : Anti-KAT3B / p300 antibody [EPR23753-55] (ab259330) at 1/1000 dilution
Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
Lane 2 : K562 (human chronic myelogenous leukemia lymphoblast), whole cell lysate
Lane 3 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate
Lane 4 : PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate
Lysates/proteins at 10 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
Fresh lysates were used in this WB.
The ~100KDa signal is a non-target band.
Lane 2 of the blot was developed using a higher-sensitivity ECL substrate.
Exposure time: 3 minutes
-
Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 cells labelling KAT3B / p300 with ab259330 at 1/500 (0.926 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green). Confocal image showing nuclear staining in NIH/3T3 cells is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/ml dilution.
-
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling KAT3B / p300 with ab259330 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.
-
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (Mouse embryonic fibroblast) cells labelling KAT3B / p300 with ab259330 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.
Datasheets and documents
-
SDS download
-
Datasheet download
Certificate of Compliance
References (0)
ab259330 has not yet been referenced specifically in any publications.