Recombinant
RabMAb

Recombinant Anti-GRB2 antibody [Y301] (ab32111)

Reviews (1)Q&A (2)References (1)

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

    Suitable for: WB, IHC-P, ICC/IF, Flow Cytmore details
    Unsuitable for: IP

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

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

  • Positive control

    • Human breast carcinoma, PC12 cells WB: 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

Applications

Our Abpromise guarantee covers the use of ab32111 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
WB 1/1000 - 1/5000. Predicted molecular weight: 25 kDa.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF 1/100 - 1/250.
Flow Cyt 1/40.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
  • Application notes
    Is unsuitable for IP.

    • Target

    • Function

      Adapter protein that provides a critical link between cell surface growth factor receptors and the Ras signaling pathway.
      Isoform GRB3-3 does not bind to phosphorylated epidermal growth factor receptor (EGFR) but inhibits EGF-induced transactivation of a RAS-responsive element. Isoform GRB3-3 acts as a dominant negative protein over GRB2 and by suppressing proliferative signals, may trigger active programmed cell death.

    • Sequence similarities

      Belongs to the GRB2/sem-5/DRK family.
      Contains 1 SH2 domain.
      Contains 2 SH3 domains.

    • Domain

      The SH3 domains mediate interaction with RALGPS1 and SHB.

    • Cellular localization

      Golgi apparatus.
    • Information by UniProt

    • Database links

      see all

    • Alternative names

      • Abundant SRC homology antibody
      • Adapter protein GRB2 antibody
      • ASH antibody
      • Ash protein antibody
      • EGFRBP GRB2 antibody
      • Epidermal growth factor receptor binding protein antibody
      • Epidermal growth factor receptor binding protein GRB2 antibody
      • GRB 2 antibody
      • GRB2 adapter protein antibody
      • Grb2 antibody
      • GRB2_HUMAN antibody
      • Grb3 3 antibody
      • Growth factor receptor bound protein 2 antibody
      • Growth factor receptor bound protein 3 antibody
      • Growth factor receptor-bound protein 2 antibody
      • HT027 antibody
      • MST084 antibody
      • MSTP084 antibody
      • NCKAP2 antibody
      • OTTHUMP00000166096 antibody
      • OTTHUMP00000166097 antibody
      • OTTHUMP00000166098 antibody
      • Protein ash antibody
      • SEM5 antibody
      • SH2/SH3 adapter GRB2 antibody
      see all

    Images

    • 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.

    • 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.

    References

    This product has been referenced in:

    • Parodi B  et al. Fumarates modulate microglia activation through a novel HCAR2 signaling pathway and rescue synaptic dysregulation in inflamed CNS. Acta Neuropathol 130:279-95 (2015). WB . Read more (PubMed: 25920452) »
    See 1 Publication for this product

    Publishing research using ab32111? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-3 of 3 Abreviews or Q&A

    Question

    What is the recommended antigen retrieval buffer for these three antibodies?

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    Answer

    Thank you for contacting us.

    The following standard heat-mediated citrate buffer antigen retrieval was used for the IHC validation of ab32111 and ab111031. Antibody ab111586 used citrate buffer in a pressure cooker at 120C for 5 minutes but a steamer or waterbath at 95-100C will be an acceptable substitute.

    Place slides in a container of 0.01 M sodium citrate buffer, pH 6.0 heated to 100°C and maintain this temperature for 20 minutes by placing the container in a steamer or heated waterbath. After 20 minutes, remove from heat and let stand at room temperature in buffer for 20 minutes. Rinse in 1X TBS with Tween (TBST) for 1 minute at RT.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Question

    What was the antigen retrieval method that was used to generate the picture on your datasheet?

    Read More
    Answer

    We used a heat mediated antigen retrieval method with sodium citrate, pH. 6.0.

    Read More

    Western blot abreview for Anti-GRB2 antibody [Y301]

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Raji)
    Loading amount
    40 µg
    Specification
    Raji
    Gel Running Conditions
    Reduced Denaturing (12%)
    Blocking step
    Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Read More

    Abcam user community

    Verified customer

    Submitted Sep 11 2009

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