Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-KAT3B / p300 antibody [EPR23495-268] - ChIP Grade – BSA and Azide free (ab275388)

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 – BSA and Azide free
  • Suitable for: ChIP, WB, ICC, Flow Cyt, IP
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-KAT3B / p300 antibody [EPR23495-268] - ChIP Grade – BSA and Azide free
    See all KAT3B / p300 primary antibodies

  • Description

    Rabbit monoclonal [EPR23495-268] to KAT3B / p300 - ChIP Grade – BSA and Azide free

  • Host species

    Rabbit

  • Specificity

    Anti-KAT3B / p300 antibody [EPR23495-268] reacts weakly with CREBBP.

  • Tested Applications & Species

    Application Species
    ChIP
    Human
    Flow Cyt
    Mouse
    ICC
    Human
    IP
    Human
    WB
    Recombinant fragment
    See all applications and species data

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: K-562, RAW 264.7 and PC-12 whole cell lysates; His-tagged human KAT3B/p300 recombinant protein. ICC: HeLa and NIH/3T3 cells. Flow cyt: NIH/3T3 cells. IP: K-562 whole cell lysate. ChIP: Chromatin prepared from K-562 cells.

  • General notes

    ab275388 is the carrier-free version of ab275378. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab275388 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    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
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Images

  • Immunoprecipitation - Anti-KAT3B / p300 antibody [EPR23495-268] - BSA and Azide free (ab275388)
    Immunoprecipitation - Anti-KAT3B / p300 antibody [EPR23495-268] - BSA and Azide free (ab275388)

    This data was developed using ab275378, the same antibody clone in a different buffer formulation.

    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.

  • ChIP - Anti-KAT3B / p300 antibody [EPR23495-268] - BSA and Azide free (ab275388)
    ChIP - Anti-KAT3B / p300 antibody [EPR23495-268] - BSA and Azide free (ab275388)

    This data was developed using ab275378, the same antibody clone in a different buffer formulation. 

    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

  • Flow Cytometry - Anti-KAT3B / p300 antibody [EPR23495-268] - BSA and Azide free (ab275388)
    Flow Cytometry - Anti-KAT3B / p300 antibody [EPR23495-268] - BSA and Azide free (ab275388)

    This data was developed using ab275378, the same antibody clone in a different buffer formulation.

    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.

  • Immunocytochemistry - Anti-KAT3B / p300 antibody [EPR23495-268] - ChIP Grade – BSA and Azide free (ab275388)
    Immunocytochemistry - Anti-KAT3B / p300 antibody [EPR23495-268] - ChIP Grade – BSA and Azide free (ab275388)

    This data was developed using ab275378, the same antibody clone in a different buffer formulation.

    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 weak 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.

  • Western blot - Anti-KAT3B / p300 antibody [EPR23495-268] - BSA and Azide free (ab275388)
    Western blot - Anti-KAT3B / p300 antibody [EPR23495-268] - BSA and Azide free (ab275388)
    All lanes : Anti-KAT3B / p300 antibody [EPR23495-268] - ChIP Grade (ab275378) at 1/1000 dilution

    Lane 1 : His-tagged human KAT3B/p300 recombinant protein (aa 1287-1666) 20 ng
    Lane 2 : His-tagged human CREBBP recombinant protein, 10 µl

    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?



    This data was developed using ab275378, the same antibody clone in a different buffer formulation. 

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    Both recombinant proteins were expressed from a E.coil expression system. Sample loaded onto lane2 was E.coil extract containing CREBBP recombinant protein.

    This product has weak cross reaction with CREBBP protein.

    Exposure time: 5.5 seconds.

  • Immunocytochemistry - Anti-KAT3B / p300 antibody [EPR23495-268] - ChIP Grade – BSA and Azide free (ab275388)
    Immunocytochemistry - Anti-KAT3B / p300 antibody [EPR23495-268] - ChIP Grade – BSA and Azide free (ab275388)

    This data was developed using ab275378, the same antibody clone in a different buffer formulation.

    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 weak 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.

  • Western blot - Anti-KAT3B / p300 antibody [EPR23495-268] - BSA and Azide free (ab275388)
    Western blot - Anti-KAT3B / p300 antibody [EPR23495-268] - BSA and Azide free (ab275388)
    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?



    This data was developed using ab275378, the same antibody clone in a different buffer formulation. 

    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.

  • Anti-KAT3B / p300 antibody [EPR23495-268] - ChIP Grade – BSA and Azide free (ab275388)
    Anti-KAT3B / p300 antibody [EPR23495-268] - ChIP Grade – BSA and Azide free (ab275388)

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