Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-RAP1GAP antibody [Y134] - BSA and Azide free (ab247250)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [Y134] to RAP1GAP - BSA and Azide free
  • Suitable for: IHC-P, ICC, WB, Flow Cyt, IP
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-RAP1GAP antibody [Y134] - BSA and Azide free
    See all RAP1GAP primary antibodies

  • Description

    Rabbit monoclonal [Y134] to RAP1GAP - BSA and Azide free

  • Host species

    Rabbit

  • Specificity

    This antibody recognises Rap1 GTPase-activating protein 1.

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    IHC-P
    Human
    IP
    Rat
    WB
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
    (Peptide available as ab139973)

  • General notes

    Ab247250 is the carrier-free version of ab32373. 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

    ab247250 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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

  • Western blot - Anti-RAP1GAP antibody [Y134] - BSA and Azide free (ab247250)
    Western blot - Anti-RAP1GAP antibody [Y134] - BSA and Azide free (ab247250)
    All lanes : Anti-RAP1GAP antibody [Y134] (ab32373) at 1/10000 dilution (purified)

    Lane 1 : Mouse brain lysate
    Lane 2 : Rat brain lysate

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size: 95 kDa
    Observed band size: 95 kDa



    This data was developed using ab32373, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-RAP1GAP antibody [Y134] - BSA and Azide free (ab247250)
    Western blot - Anti-RAP1GAP antibody [Y134] - BSA and Azide free (ab247250)
    Anti-RAP1GAP antibody [Y134] (ab32373) at 1/10000 dilution (purified) + Jurkat cell lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 95 kDa
    Observed band size: 95 kDa



    This data was developed using ab32373, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-RAP1GAP antibody [Y134] - BSA and Azide free (ab247250)
    Western blot - Anti-RAP1GAP antibody [Y134] - BSA and Azide free (ab247250)
    Anti-RAP1GAP antibody [Y134] (ab32373) at 1/10000 dilution (purified) + SH-SY5Y cell lysate at 20 µg

    Secondary
    Anti-rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 95 kDa
    Observed band size: 95 kDa



    This data was developed using ab32373, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-RAP1GAP antibody [Y134] - BSA and Azide free (ab247250)
    Western blot - Anti-RAP1GAP antibody [Y134] - BSA and Azide free (ab247250)
    Anti-RAP1GAP antibody [Y134] (ab32373) at 1/50000 dilution (unpurified) + Jurkat cell lysate

    Predicted band size: 95 kDa
    Observed band size: 95 kDa



    This data was developed using ab32373, the same antibody clone in a different buffer formulation.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAP1GAP antibody [Y134] - BSA and Azide free (ab247250)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAP1GAP antibody [Y134] - BSA and Azide free (ab247250)

    This data was developed using ab32373, the same antibody clone in a different buffer formulation.

    Immunohistochemical staining of paraffin embedded human cerebral cortex with purified ab32373 at a working dilution of 1/1000. The secondary antibody used is ab97051, a goat anti-rabbit IgG (H&L) at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAP1GAP antibody [Y134] - BSA and Azide free (ab247250)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAP1GAP antibody [Y134] - BSA and Azide free (ab247250)

    This data was developed using ab32373, the same antibody clone in a different buffer formulation.

    Immunohistochemical staining of paraffin-embedded human breast cancer tissue using unpurified ab32373 at 1/100 dilution.
  • Immunocytochemistry - Anti-RAP1GAP antibody [Y134] - BSA and Azide free (ab247250)
    Immunocytochemistry - Anti-RAP1GAP antibody [Y134] - BSA and Azide free (ab247250)
    This data was developed using ab32373, the same antibody clone in a different buffer formulation.Immunofluorescence staining of HeLa cells with purified ab32373 at a working dilution of 1/250, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4 % PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab32373 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.
  • Immunocytochemistry - Anti-RAP1GAP antibody [Y134] - BSA and Azide free (ab247250)
    Immunocytochemistry - Anti-RAP1GAP antibody [Y134] - BSA and Azide free (ab247250)
    This data was developed using ab32373, the same antibody clone in a different buffer formulation.ICC/IF image of unpurified ab32373 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab32373, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h.Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Flow Cytometry - Anti-RAP1GAP antibody [Y134] - BSA and Azide free (ab247250)
    Flow Cytometry - Anti-RAP1GAP antibody [Y134] - BSA and Azide free (ab247250)
    This data was developed using ab32373, the same antibody clone in a different buffer formulation.Overlay histogram showing Jurkat cells fixed in 4% PFA and stained with purified ab32373 at a dilution of 1/30 (red line). The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit at a dilution of 1/500. Rabbit monoclonal IgG was used as an isotype control (black line) and cells incubated in the absence of both primary and secondary antibody were used as a negative control (blue line).
  • Flow Cytometry - Anti-RAP1GAP antibody [Y134] - BSA and Azide free (ab247250)
    Flow Cytometry - Anti-RAP1GAP antibody [Y134] - BSA and Azide free (ab247250)
    This data was developed using ab32373, the same antibody clone in a different buffer formulation.Overlay histogram showing SH-SY5Y cells stained with unpurified ab32373 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32373, 1/100) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • Immunoprecipitation - Anti-RAP1GAP antibody [Y134] - BSA and Azide free (ab247250)
    Immunoprecipitation - Anti-RAP1GAP antibody [Y134] - BSA and Azide free (ab247250)
    This data was developed using ab32373, the same antibody clone in a different buffer formulation.ab32373 (purified) at 1/20 immunoprecipitating RAP1GAP in 10 μg C6 cell lysate (Lanes 1 and 2, observed at 82-95 kDa). Lane 3 - Rabbit monoclonal IgG (ab172730). For western blotting, HRP Veriblot for IP Detection Reagent (ab131366) was used for detection (1/1000). Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST
  • Anti-RAP1GAP antibody [Y134] - BSA and Azide free (ab247250)
    Anti-RAP1GAP antibody [Y134] - BSA and Azide free (ab247250)

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