Overview

  • Product name

    Anti-RbAp48 antibody [EPR3411]
    See all RbAp48 primary antibodies
  • Description

    Rabbit monoclonal [EPR3411] to RbAp48
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, WB, IP, IHC-P, Flow Cytmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide. within Human RbAp48 aa 1-100 (N terminal). The exact sequence is proprietary.
    Database link: Q09028

  • Positive control

    • WB: HeLa, Jurkat, NIH/3T3, 293 and HeLa, whole cell lysates., Mouse spleen, Rat spleen and Human fetal spleen tissue lysates IHC-P:Human breast cancer, rat pancreas and Mouse cerebrum FC: MCF7 cells IP: HeLa cells ICC/IF: MCF-7 and HeLa cells.
  • General notes

     

     

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab79416 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
ICC/IF 1/100 - 1/250.
WB 1/10000 - 1/50000. Detects a band of approximately 48 kDa (predicted molecular weight: 48 kDa).
IP 1/20.

For unpurified use at 1/50

IHC-P 1/8000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified use at 1/100 - 1/250

Flow Cyt 1/20.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

For unpurified use at 1/50

Target

  • Function

    Core histone-binding subunit that may target chromatin assembly factors, chromatin remodeling factors and histone deacetylases to their histone substrates in a manner that is regulated by nucleosomal DNA. Component of several complexes which regulate chromatin metabolism. These include the chromatin assembly factor 1 (CAF-1) complex, which is required for chromatin assembly following DNA replication and DNA repair; the core histone deacetylase (HDAC) complex, which promotes histone deacetylation and consequent transcriptional repression; the nucleosome remodeling and histone deacetylase complex (the NuRD complex), which promotes transcriptional repression by histone deacetylation and nucleosome remodeling; the PRC2/EED-EZH2 complex, which promotes repression of homeotic genes during development; and the NURF (nucleosome remodeling factor) complex.
  • Sequence similarities

    Belongs to the WD repeat RBAP46/RBAP48/MSI1 family.
    Contains 6 WD repeats.
  • Cellular localization

    Nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • CAF I p48 antibody
    • CAF-1 subunit C antibody
    • CAF-I 48 kDa subunit antibody
    • CAF-I p48 antibody
    • Chromatin assembly factor 1 subunit C antibody
    • Chromatin assembly factor I p48 subunit antibody
    • Chromatin assembly factor/CAF 1 p48 subunit antibody
    • Histone-binding protein RBBP4 antibody
    • MSI1 protein homolog antibody
    • Nucleosome-remodeling factor subunit RBAP48 antibody
    • NURF55 antibody
    • RB binding protein 4 chromatin remodeling factor antibody
    • RbAp 48 antibody
    • RBAP48 antibody
    • RBBP-4 antibody
    • RBBP4 antibody
    • RBBP4_HUMAN antibody
    • Retinoblastoma binding protein 4 antibody
    • Retinoblastoma binding protein p48 antibody
    • Retinoblastoma-binding protein 4 antibody
    • Retinoblastoma-binding protein p48 antibody
    see all

Images

  • All lanes : Anti-RbAp48 antibody [EPR3411] (ab79416) at 0.006 µg/ml (purified)

    Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
    Lane 2 : Mouse spleen lysates
    Lane 3 : Rat spleen lysates

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 48 kDa



    Blocking and diluting buffer: 5% NFDM/TBST. 

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat pancreas tissue sections labeling RbAp48 with Purified ab79416 at 1:8000 dilution (0.014 µg/ml). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0)ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody.Negative control:PBS instead of the primary antibody.Hematoxylinwas used as a counterstain
  • ab79416 (purified) at 1:20 dilution (2µg) immunoprecipitating RbAp48 in HeLa whole cell lysate.
    Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10µg
    Lane 2 (+): ab79416 & HeLa whole cell lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab79416 in HeLa whole cell lysate
    For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
    Blocking and diluting buffer: 5% NFDM/TBST.

  • Flow Cytometry analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling RbAp48 with purified ab79416 at 1:20 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse cerebrum tissue sections labeling RbAp48 with Purified ab79416 at 1:8000 dilution (0.014 µg/ml). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0)ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody.Negative control:PBS instead of the primary antibody.Hematoxylinwas used as a counterstain
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast cancer tissue sections labeling RbAp48 with Purified ab79416 at 1:8000 dilution (0.014 µg/ml). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0)ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody.Negative control:PBS instead of the primary antibody.Hematoxylinwas used as a counterstain
  • Anti-RbAp48 antibody [EPR3411] (ab79416) at 0.006 µg/ml (purified) + Human fetal spleen lysates at 15 µg

    Secondary
    Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

    Predicted band size: 48 kDa



    Blocking and diluting buffer: 5% NFDM/TBST. 

  • ab79416 staining RbAp48 in MCF-7 (human breast carcinoma) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Samples were incubated with primary antibody at a dilution of 1/500. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a dilution of 1/1000. DAPI was used as a nuclear counterstain.

    Negative control 1: PBS only.

  • All lanes : Anti-RbAp48 antibody [EPR3411] (ab79416) at 1/50000 dilution (Unpurifed)

    Lane 1 : HeLa lysate
    Lane 2 : Jurkat lysate
    Lane 3 : NIIH/3T3 lysate
    Lane 4 : 293 lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

    Predicted band size: 48 kDa
    Observed band size: 48 kDa

  • Unpurifed ab79416, at 1/100 dilution, staining RbAp48 in human breast carcinoma tissue by Immunohistochemistry.

  • Immunofluorescent staining of RbAp48 in HeLa cells using unpurified ab79416 at 1/100 dilution.

References

This product has been referenced in:

  • Patel D  et al. The histone demethylase LSD1 regulates inner ear progenitor differentiation through interactions with Pax2 and the NuRD repressor complex. PLoS One 13:e0191689 (2018). Read more (PubMed: 29370269) »
  • Kraushaar DC  et al. The gene repressor complex NuRD interacts with the histone variant H3.3 at promoters of active genes. Genome Res 28:1646-1655 (2018). Read more (PubMed: 30254051) »
See all 9 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Answer

Thank you very much for your inquiry. As x is out of the office this week, I have follow up in regards to your question with the laboratory.

The laboratory has confirmed that the immunogen shares only 64% identity with RbAp46 protein, sowe would not predict cross-reactivity. We expect this antibodytherefore to bespecific for RbAp48, however we have not tested this experimentally.

The information you are referring to on the datasheet, means that the antibody belongs to that family of proteins. The protein RbAp48 is however a distinct protein, sharing some homology with the family members. Please see also the SwissProt entry of RbAp48 for additional information: http://www.uniprot.org/uniprot/Q09028

I hope this information is helpful . Please do not hesitate to get back to us, should you or the customer have anyquestions or concerns.

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Answer


The choice is perfect. Both ab79416 and ab109285are guaranteed to work in WB, IP, IHC-P and in Mouse.

Read More

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