Recombinant Anti-GRB2 antibody [Y237] (ab32037)
![Western blot - Anti-GRB2 antibody [Y237] (ab32037)](https://www.abcam.com/ps/products/32/ab32037/Images/ab32037-459633-anti-grb2-antibody-y237-western-blot-wildtype-hela-grb2-knockout-hela-hl60-caco2.jpg "Western blot - Anti-GRB2 antibody [Y237] (ab32037)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [Y237] to GRB2
- Suitable for: WB, Flow Cyt (Intra), IP, IHC-P, ICC/IF
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-GRB2 antibody [Y237]
See all GRB2 primary antibodies -
Description
Rabbit monoclonal [Y237] to GRB2 -
Host species
Rabbit -
Tested applications
Suitable for: WB, Flow Cyt (Intra), IP, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide within Human GRB2 aa 200 to the C-terminus (C terminal). The exact sequence is proprietary.
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Positive control
- WB: HeLa, HL-60, Caco-2, HEK-293, PC-12 and NIH/3T3 cell lysates. IHC-P: Human ovarian carcinoma tissue. ICC/IF: HeLa cells. Flow Cyt (intra): HeLa cells.
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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
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
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
Y237 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab32037 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 | (2) |
1/5000 - 1/10000. Detects a band of approximately 25 kDa (predicted molecular weight: 25 kDa).
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| Flow Cyt (Intra) |
1/20.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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| IP |
Use at an assay dependent concentration.
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| IHC-P |
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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| ICC/IF | (1) |
1/50 - 1/100.
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| Notes |
|---|
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WB
1/5000 - 1/10000. Detects a band of approximately 25 kDa (predicted molecular weight: 25 kDa). |
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Flow Cyt (Intra)
1/20. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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IP
Use at an assay dependent concentration. |
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IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
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ICC/IF
1/50 - 1/100. |
Target
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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
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Database links
- Entrez Gene: 2885 Human
- Entrez Gene: 14784 Mouse
- Entrez Gene: 81504 Rat
- Omim: 108355 Human
- SwissProt: P62993 Human
- SwissProt: Q60631 Mouse
- SwissProt: P62994 Rat
- Unigene: 444356 Human
see all -
Alternative names
- Abundant SRC homology antibody
- Adapter protein GRB2 antibody
- ASH antibody
see all
Images
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Western blot - Anti-GRB2 antibody [Y237] (ab32037)All lanes : Anti-GRB2 antibody [Y237] (ab32037) at 1/5000 dilution
Lane 1 : Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : GRB2 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 3 : HL-60 (Human promyelocytic leukemia cell line) whole cell lysate
Lane 4 : Caco-2 (Human colorectal adenocarcinoma cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 25 kDa
Observed band size: 25 kDaLanes 1 - 4: Merged signal (red and green). Green - ab32037 observed at 25 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
ab32037 was shown to react with GRB2 in western blot. The band observed in the CRISPR-Cas9 edited cell lysate lane above 25 kDa is likely to represent an insertion. This has not been investigated further and the functional properties of the gene product have not been determined. Cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab32037 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 5000 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 h at room temperature before imaging.
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![Immunocytochemistry/ Immunofluorescence - Anti-GRB2 antibody [Y237] (ab32037)](https://www.abcam.com/ps/products/32/ab32037/Images/ab32037-349908-anti-grb2-antibody-Y237-immunohistochemistry-heLa-human.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-GRB2 antibody [Y237] (ab32037)Immunocytochemistry/immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% TritonX-100 permeabilised HeLa (Human cervix adenocarcinoma epithelial cell) cells staining GRB2 with ab32037 at 1/200 dilution (green), followed by AlexaFluor®488 Goat anti-Rabbit secondary antibody (ab150077) at 1/1000 dilution (blue). Confocal image showing nuclear and weak cytoplasmic staining in HeLa cell line. Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) was used as a counterstain at 1/200 dilution (red). DAPI was used as a nuclear counterstain.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is [secondary antibody name] ab150077 at 1/1000 dilution.Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRB2 antibody [Y237] (ab32037)](https://www.abcam.com/ps/products/32/ab32037/Images/ab32037-43704-anti-grb2-antibody-y237-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRB2 antibody [Y237] (ab32037)Immunohistochemical analysis of GRB2 expression in paraffin embedded human ovarian carcinoma, using 1/100 ab32037.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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![Flow Cytometry (Intracellular) - Anti-GRB2 antibody [Y237] (ab32037)](https://www.abcam.com/ps/products/32/ab32037/Images/ab32037-287292-anti-grb2-antibody-y237-flow-cytometry.jpg)
Flow Cytometry (Intracellular) - Anti-GRB2 antibody [Y237] (ab32037)Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling GRB2 (red) with ab32037 at a 1/20 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluorr® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (Black) (ab172730). Blue (unlabeled control) - Cell without incubation with primary antibody and secondary antibody (Blue).
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![Western blot - Anti-GRB2 antibody [Y237] (ab32037)](https://www.abcam.com/ps/products/32/ab32037/Images/ab32037-257525-anti-grb2-antibody-y237-western-blot.jpg)
Western blot - Anti-GRB2 antibody [Y237] (ab32037)All lanes : Anti-GRB2 antibody [Y237] (ab32037) at 1/10000 dilution
Lane 1 : PC-12 (rat adrenal gland pheochromocytoma) cell lysate
Lane 2 : NIH/3T3 (mouse embryonic fibroblast) cell lysate
Lane 3 : HEK293 (human epithelial cell line from embryonic kidney) cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/2000 dilution
Predicted band size: 25 kDa
Observed band size: 25 kDaBlocking/Dilution buffer : 5% NFDM/TBST
Exposure time : 3 minutes
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![Western blot - Anti-GRB2 antibody [Y237] (ab32037)](https://www.abcam.com/ps/products/32/ab32037/Images/ab32037-43702-anti-grb2-antibody-y237-western-blot.jpg)
Western blot - Anti-GRB2 antibody [Y237] (ab32037)This image is a courtesy of Anonymous AbreviewAnti-GRB2 antibody [Y237] (ab32037) at 1/5000 dilution + Lysate prepared from human Mesothelial cell line MeT-5A at 4 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/2000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 25 kDa
Observed band size: 24 kDa why is the actual band size different from the predicted?
Exposure time: 1 minute
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![Western blot - Anti-GRB2 antibody [Y237] (ab32037)](https://www.abcam.com/ps/products/32/ab32037/Images/ab32037-43701-anti-grb2-antibody-y237-western-blot.jpg)
Western blot - Anti-GRB2 antibody [Y237] (ab32037)Anti-GRB2 antibody [Y237] (ab32037) at 1/5000 dilution +Recombinant Human GRB2 protein (ab85917) at 0.01 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 25 kDa
Exposure time: 2 minutes -
![Anti-GRB2 antibody [Y237] (ab32037)](https://www.abcam.com/ps/products/32/ab32037/Images/ab32037-10-benefits-of-recombinant-antibodies.png)
Anti-GRB2 antibody [Y237] (ab32037)
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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Datasheet download
References (19)
ab32037 has been referenced in 19 publications.
- Wang J et al. Grb2 Induces Cardiorenal Syndrome Type 3: Roles of IL-6, Cardiomyocyte Bioenergetics, and Akt/mTOR Pathway. Front Cell Dev Biol 9:630412 (2021). PubMed: 33829014
- Wang Y et al. Inhibition of bone morphogenetic protein receptor 2 suppresses pancreatic ductal adenocarcinoma growth by regulating GRB2/PI3K/AKT axis. Ann Transl Med 9:557 (2021). PubMed: 33987255
- Lewis TR et al. NHD2-15, a novel antagonist of Growth Factor Receptor-Bound Protein-2 (GRB2), inhibits leukemic proliferation. PLoS One 15:e0236839 (2020). PubMed: 32780746
- Sun C et al. Long noncoding RNA AC092171.4 promotes hepatocellular carcinoma progression by sponging microRNA-1271 and upregulating GRB2. Aging (Albany NY) 12:14141-14156 (2020). PubMed: 32692718
- Yang Y et al. Human cytomegalovirus IE2 protein regulates macrophage-mediated immune escape by upregulating GRB2 expression in UL122 genetically modified mice. Biosci Trends 13:502-509 (2020). PubMed: 31866613
