Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (ab185971)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR13059(2)] to RYBP - ChIP Grade
  • Suitable for: ChIP, IP, Flow Cyt, ICC/IF, IHC-P, WB
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-RYBP antibody [EPR13059(2)] - ChIP Grade
    See all RYBP primary antibodies

  • Description

    Rabbit monoclonal [EPR13059(2)] to RYBP - ChIP Grade

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    ChIP
    Mouse
    Human
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Human
    IP
    Mouse
    Human
    WB
    Rat
    Human
    See all applications and species data

  • Immunogen

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

  • Positive control

    • K562, SW480, HepG2 and rat PC12 cell lysates; Human placenta and Human clear carcinoma of kidney tissues; HepG2, SW480 and K562 cells IP: NIH/3T3 cell lysates Flow Cyt: A431 cells

  • 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
    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.

Properties

Images

  • Western blot - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (ab185971)
    Western blot - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (ab185971)
    All lanes : Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (ab185971) at 1/1000 dilution

    Lane 1 : K562 cell lysate
    Lane 2 : SW480 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : goat anti-rabbit IgG, (H+L), peroxidase conjugated at 1/1000 dilution

    Developed using the ECL technique.

    Predicted band size: 25 kDa
    Observed band size: 32 kDa
    why is the actual band size different from the predicted?

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (ab185971)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (ab185971)

    Immunohistochemical analysis of paraffin-embedded, Human clear cell carcinoma kidney tissue labeling RYBP with ab185971 at a 1/100 dilution. Counter stained with hematoxylin.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (ab185971)
    Immunocytochemistry/ Immunofluorescence - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (ab185971)

    Immunofluorescence analysis of, paraformaldehyde-fixed, HepG2 cells labeling RYBP with ab185971 at a 1/250 dilution. As secondary antibody goat anti-rabbit IgG (Alexa Fluor®555) was used at a 1/200. In blue DAPI staining.

  • Immunoprecipitation - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (ab185971)
    Immunoprecipitation - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (ab185971)

    ab185971 (purified) at 1/20 dilution (11.7µg/mL) immunoprecipitating RYBP in NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate 10 µg.

    Lane 1 (input): NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate 10 µg
    Lane 2 (+): ab185971 & NIH/3T3 whole cell lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab185971 in NIH/3T3 whole cell lysate

    For western blotting, ab185971 at 1/500 dilution (0.468 µg/mL) and veriBlot for IP secondary antibody (HRP) (ab131366) at 1/1000 dilution was used.

    Blocking and diluting buffer: 5% NFDM /TBST.

  • Flow Cytometry - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (ab185971)
    Flow Cytometry - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (ab185971)

    Flow Cytometry analysis of A431 (Human epidermoid carcinoma epithelial cell) cells labeling RYBP with purified ab185971 at 1/20 dilution (11.7 µg/mL) (Red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) (black). Unlabeled control - Unlabelled cells (blue).

  • ChIP - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (ab185971)
    ChIP - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (ab185971)
    Chromatin was prepared from NIH/3T3 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 ab185971 (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%20protocol
  • Flow Cytometry - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (ab185971)
    Flow Cytometry - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (ab185971)

    Flow cytometry analysis of paraformaldehyde-fixed K562 cells labeling RYBP with ab185971 at a 1/40 dilution and secondary antibody goat anti-rabbit IgG (FITC, red) at a 1/150 dilution, or negative control rabbit IgG (green).

  • ChIP - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (ab185971)
    ChIP - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (ab185971)
    Chromatin was prepared from HEK293 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 ab185971 (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%20protocol
  • Western blot - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (ab185971)
    Western blot - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (ab185971)
    Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (ab185971) at 1/10000 dilution + HepG2 cell lysate at 20 µg

    Secondary
    goat anti-rabbit IgG, (H+L), peroxidase conjugated at 1/1000 dilution

    Developed using the ECL technique.

    Predicted band size: 25 kDa
    Observed band size: 32 kDa why is the actual band size different from the predicted?

  • Western blot - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (ab185971)
    Western blot - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (ab185971)
    Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (ab185971) at 1/1000 dilution + rat PC12 cell lysate at 10 µg

    Secondary
    goat anti-rabbit IgG, (H+L), peroxidase conjugated at 1/1000 dilution

    Developed using the ECL technique.

    Predicted band size: 25 kDa
    Observed band size: 32 kDa why is the actual band size different from the predicted?

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (ab185971)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (ab185971)

    Immunohistochemical analysis of paraffin-embedded, Human placenta tissue labeling RYBP with ab185971 at a 1/100 dilution. Counter stained with hematoxylin.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (ab185971)
    Immunocytochemistry/ Immunofluorescence - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (ab185971)

    Immunofluorescence analysis of, paraformaldehyde-fixed, SW480 cells labeling RYBP with ab185971 at a 1/250 dilution. As secondary antibody goat anti-rabbit IgG (Alexa Fluor®555) was used at a 1/200. In blue DAPI staining.

  • Immunoprecipitation - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (ab185971)
    Immunoprecipitation - Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (ab185971)

    Western blot analysis on immunoprecipitation pellet from HepG2 cell lysate labeling RYBP with ab185971 at 1/50 dilution and goat anti-rabbit IgG, (H+L), peroxidase conjugated at a 1/1000 dilution.

  • Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (ab185971)
    Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (ab185971)

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