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AB281892

Anti-GRB2 antibody [81/GRB2] - BSA and Azide free

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Mouse Recombinant Monoclonal GRB2 antibody. Carrier free. Suitable for WB and reacts with Mouse, Rat, Human samples.

View Alternative Names

ASH, GRB2, Growth factor receptor-bound protein 2, Adapter protein GRB2, Protein Ash, SH2/SH3 adapter GRB2

2 Images
Western blot - Anti-GRB2 antibody [81/GRB2] - BSA and Azide free (AB281892)
  • WB

Lab

Western blot - Anti-GRB2 antibody [81/GRB2] - BSA and Azide free (AB281892)

This data was developed using ab281846, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Exposure time : 7 seconds

All lanes:

Western blot - Anti-GRB2 antibody [81/GRB2] (<a href='/en-us/products/primary-antibodies/grb2-antibody-81-grb2-ab281846'>ab281846</a>) at 1/1000 dilution

Lane 1:

HeLa (human epithelial cell line from cervix adenocarcinoma), whole cell lysate at 20 µg

Lane 2:

A431 (human epidermoid carcinoma epithelial cell), whole cell lysate at 20 µg

Lane 3:

NIH/3T3 (mouse embryonic fibroblast), whole cell lysate at 20 µg

Lane 4:

PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate at 20 µg

Lane 5:

Human tonsil tissue lysate at 20 µg

Lane 6:

Mouse testis tissue lysate at 20 µg

Lane 7:

Rat testis tissue lysate at 20 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Predicted band size: 25 kDa

Observed band size: 26 kDa

false

Western blot - Anti-GRB2 antibody [81/GRB2] - BSA and Azide free (AB281892)
  • WB

Lab

Western blot - Anti-GRB2 antibody [81/GRB2] - BSA and Azide free (AB281892)

This data was developed using ab281846, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS

Lanes 1-4 : Merged signal (red and green). Green - ab281846 observed at 26KDa. Red - loading control ab181602 (Rabbit monoclonal [EPR16891] to GAPDH) observed at 36 kDa.

Lanes 1-2 : ab281846 Anti-GRB2 antibody was shown to react with GRB2 in HCT116 cells in Western blot. Loss of signal was observed when GRB2 knockout cell line ab273715 (knockout cell lysate ab275248) was used. Wild-type and GRB2 knockout samples were subjected to SDS-PAGE.

ab281846 and Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) were incubated at 4℃ overnight at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680CW) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800RD) preadsorbed (ab216772) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-GRB2 antibody [81/GRB2] (<a href='/en-us/products/primary-antibodies/grb2-antibody-81-grb2-ab281846'>ab281846</a>) at 1/1000 dilution

Lane 1:

Wild type HCT116 (human colorectal carcinoma epithelial cell), whole cell lysate at 20 µg

Lane 2:

GRB2 knockout HCT116 (<a href='/en-us/products/cell-lines/human-grb2-knockout-hct116-cell-line-ab273715'>ab273715</a>) whole cell lysate at 20 µg

Lane 2:

Western blot - Human GRB2 knockout HCT116 cell line (<a href='/en-us/products/cell-lines/human-grb2-knockout-hct116-cell-line-ab273715'>ab273715</a>)

Lane 3:

HeLa (human epithelial cell line from cervix adenocarcinoma), whole cell lysate at 20 µg

Lane 4:

293T (human embryonic kidney epithelial cell), whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Mouse IgG H&L (IRDye® 800CW) (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-800cw-preadsorbed-ab216772'>ab216772</a>) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-680rd-preadsorbed-ab216777'>ab216777</a>) at 1/10000 dilution

Predicted band size: 25 kDa

Observed band size: 26 kDa

false

Key facts

Host species

Mouse

Clonality

Monoclonal

Clone number

81/GRB2

Isotype

IgG1

Carrier free

Yes

Reacts with

Mouse, Human, Rat

Applications

WB

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "<p></p>", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "<p></p>" }, "Mouse": { "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "<p></p>", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "<p></p>" }, "Rat": { "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "<p></p>", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "<p></p>", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "<p></p>" } } }
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Product details

ab281892 is the carrier-free version of ab281846.

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

Want a custom formulation?
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Compatibility
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

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Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
pH: 7.2 - 7.4 Constituents: PBS
Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
Do Not Freeze
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

The GRB2 protein also known as growth factor receptor-bound protein 2 is instrumental in signal transduction. GRB2 has a molecular weight of about 25 kDa. It comprises one SH2 domain and two SH3 domains which facilitate its role in linking receptor tyrosine kinases to downstream signaling molecules. GRB2 is ubiquitously expressed in various tissues signifying its importance in diverse cellular functions.
Biological function summary

GRB2 functions as an adaptor protein that connects activated receptor tyrosine kinases to intracellular pathways. It often forms complexes with other proteins such as the SOS protein which further propagates signaling cascades critical for cell proliferation and differentiation. GRB2’s ability to mediate these interactions contributes to cellular responses to external stimuli including growth factors and hormones.

Pathways

GRB2 plays an important role in the Ras-MAPK signaling pathway. It interacts with proteins like SOS and Ras enabling signal transduction from the cell surface to the nucleus which is vital for processes such as cell growth and survival. GRB2 is also implicated in the PI3K-Akt pathway connecting it to another set of signaling proteins that regulate metabolism growth and survival.

GRB2 has significant associations with cancer and immune disorders. Aberrant activation of pathways involving GRB2 can lead to uncontrolled cell proliferation contributing to oncogenesis in different cancers. Additionally GRB2’s interactions with proteins like BCR-ABL in chronic myeloid leukemia highlight its potential role as a therapeutic target. GRB2 inhibitors could therefore offer promising avenues for treating such conditions by disrupting its pathological signaling interactions.
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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:
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Target data

Non-enzymatic adapter protein that plays a pivotal role in precisely regulated signaling cascades from cell surface receptors to cellular responses, including signaling transduction and gene expression (PubMed : 11726515, PubMed : 37626338). Thus, participates in many biological processes including regulation of innate and adaptive immunity, autophagy, DNA repair or necroptosis (PubMed : 35831301, PubMed : 37626338, PubMed : 38182563). Controls signaling complexes at the T-cell antigen receptor to facilitate the activation, differentiation, and function of T-cells (PubMed : 36864087, PubMed : 9489702). Mechanistically, engagement of the TCR leads to phosphorylation of the adapter protein LAT, which serves as docking site for GRB2 (PubMed : 9489702). In turn, GRB2 establishes a a connection with SOS1 that acts as a guanine nucleotide exchange factor and serves as a critical regulator of KRAS/RAF1 leading to MAPKs translocation to the nucleus and activation (PubMed : 12171928, PubMed : 25870599). Functions also a role in B-cell activation by amplifying Ca(2+) mobilization and activation of the ERK MAP kinase pathway upon recruitment to the phosphorylated B-cell antigen receptor (BCR) (PubMed : 25413232, PubMed : 29523808). Plays a role in switching between autophagy and programmed necrosis upstream of EGFR by interacting with components of necrosomes including RIPK1 and with autophagy regulators SQSTM1 and BECN1 (PubMed : 35831301, PubMed : 38182563). Regulates miRNA biogenesis by forming a functional ternary complex with AGO2 and DICER1 (PubMed : 37328606). Functions in the replication stress response by protecting DNA at stalled replication forks from MRE11-mediated degradation. Mechanistically, inhibits RAD51 ATPase activity to stabilize RAD51 on stalled replication forks (PubMed : 38459011). Additionally, directly recruits and later releases MRE11 at DNA damage sites during the homology-directed repair (HDR) process (PubMed : 34348893).. Isoform 2. Does not bind to phosphorylated epidermal growth factor receptor (EGFR) but inhibits EGF-induced transactivation of a RAS-responsive element. Acts as a dominant negative protein over GRB2 and by suppressing proliferative signals, may trigger active programmed cell death. Mechanistically, inhibits RAS-ERK signaling and downstream cell proliferation by competing with GRB2 for SOS1 binding and thus by regulating SOS1 membrane recruitment (PubMed : 36171279).
See full target information GRB2
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