Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-GRB2 antibody [Y301] (ab32111)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [Y301] to GRB2
  • Suitable for: WB, IHC-P, ICC/IF, Flow Cyt
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-GRB2 antibody [Y301]
    See all GRB2 primary antibodies

  • Description

    Rabbit monoclonal [Y301] to GRB2

  • Host species

    Rabbit

  • Specificity

    ab32111 recognises GRB2. This antibody can also detect the splice isoform GRB3-3 of GRB2.

  • Tested Applications & Species

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

  • Immunogen

    Synthetic peptide within Human GRB2 aa 1-100 (N terminal). The exact sequence is proprietary.

  • Positive control

    • Human breast carcinoma, PC12 cells WB: HCT116, HeLa, PC-12, NIH/3T3, HEK-293, RAW 264.7 and C6 cell lysates

  • 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-GRB2 antibody [Y301] (ab32111)
    Western blot - Anti-GRB2 antibody [Y301] (ab32111)
    All lanes : Anti-GRB2 antibody [Y301] (ab32111) at 1/2000 dilution

    Lane 1 : Wild-type HCT116 cell lysate
    Lane 2 : GRB2 knockout HCT116 cell lysate
    Lane 3 : HeLa cell lysate
    Lane 4 : HEK293 cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 25 kDa
    Observed band size: 25 kDa



    Lanes 1 - 4: Merged signal (red and green). Green - ab32111 observed at 25 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

    ab32111 was shown to react with GRB2 in HCT 116 wild-type cells in western blot with loss of signal observed in GRB2 knockout cell line ab273715 (GRB2 knockout cell lysate ab275248). HCT 116 wild-type and GRB2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab32111 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 2000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRB2 antibody [Y301] (ab32111)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRB2 antibody [Y301] (ab32111)

    Immunohistochemical analysis of GRB2 expression in paraffin embedded human breast carcinoma tissue section, using 1/100 ab32111. Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-GRB2 antibody [Y301] (ab32111)
    Immunocytochemistry/ Immunofluorescence - Anti-GRB2 antibody [Y301] (ab32111)
    Immunofluorescent analysis of GRB2 expression in PC12 cells, using 1/100 ab32111.
  • Flow Cytometry - Anti-GRB2 antibody [Y301] (ab32111)
    Flow Cytometry - Anti-GRB2 antibody [Y301] (ab32111)
    Overlay histogram showing SH-SY5Y cells stained with ab32111 (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 (ab32111, 1/100 dilution) 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.

  • Western blot - Anti-GRB2 antibody [Y301] (ab32111)
    Western blot - Anti-GRB2 antibody [Y301] (ab32111)
    All lanes : Anti-GRB2 antibody [Y301] (ab32111) at 1/5000 dilution (purified)

    Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
    Lane 2 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates
    Lane 3 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates
    Lane 4 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates
    Lane 5 : RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates
    Lane 6 : C6 (Rat glial tumor glial cell) whole cell 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: 25 kDa
    Observed band size: 25 kDa



    Blocking/Diluting Buffer and concentration: 5% NFDM/TBST

    We are unsure how to define the extra bands.

  • Anti-GRB2 antibody [Y301] (ab32111)
    Anti-GRB2 antibody [Y301] (ab32111)

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