Invitrogen

GRB2 Polyclonal Antibody

Advanced Verification

This Antibody was verified by Independent antibody to ensure that the antibody binds to the antigen stated.

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Datasheet

Protocols

Questions & Answers

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Datasheet

Protocols

Questions & Answers

Tech Support

Product Image Gallery

GRB2 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of GRB2 was performed using 70% confluent log phase A-431 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with GRB2 Rabbit Polyclonal Antibody(Product # PA5-17692) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

GRB2 Antibody in Western Blot (WB) — Western blot analysis was performed on membrane enriched extracts (30 µg lysate) of MCF7 (Lane 1), A-431 (Lane 2), H1975 (Lane 3), PC-3 (Lane 4), DU-145 (Lane 5), MDA-MB-231 (Lane 6), Ramos (Lane 7), HEK-293 (Lane 8), 3T3-L1 (Lane 9), Jurkat (Lane 10) and A549 (Lane 11). The blots were probed with Rabbit Anti-GRB2 Polyclonal Antibody (Product # PA5-17692, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 25 kDa band corresponding to GRB2 was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0342BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane using iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

GRB2 Antibody — Antibody specificity was demonstrated by CRISPR-Cas9 mediated knockout of target protein. A loss of signal was observed for target protein in HeLa Cas9 cell line transduced with GRB2 Lentiviral sgRNA compared to control cell line using Anti-GRB2 Polyclonal Antibody (Product # PA5-17692).

Product Details

PA5-17692

Applications

Tested Dilution

Publications

Western Blot (WB)

1:1,000

Immunocytochemistry (ICC/IF)

1:100

Product Specifications

Species Reactivity

Bovine, Hamster, Human, Mouse, Non-human primate, Rat

Host/Isotype

Rabbit / IgG

Class

Polyclonal

Type

Antibody

Immunogen

Synthetic peptide corresponding to carboxy-terminal residues of human GRB2

View immunogen

Conjugate

Unconjugated

Form

Liquid

Concentration

217 µg/mL

Purification

Antigen affinity chromatography

Storage buffer

0.01M HEPES, pH 7.5, with 0.15M NaCl, 100µg/mL BSA, 50% glycerol

Contains

no preservative

Storage conditions

-20°C

RRID

AB_10986571

Product Specific Information

It is not recommended to aliquot this antibody.

Target Information

GRB 2 protein contains one SH2 domain flanked on both sides by an SH3 domain. In response to EGF, GRB 2 binds, through its SH3 domains, to the guanine nucleotide exchange factor, Sos. The GRB 2-Sos complex then binds the EGF receptor. These interactions play a critical role in the activation of the p21 ras signaling pathway.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

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Bioinformatics

Protein Aliases: abundant SRC homology; Adapter protein GRB2; ASH protein; epidermal growth factor receptor-binding protein GRB2; Growth factor receptor-bound protein 2; growth factor receptor-bound protein 3; HT027; OTTHUMP00000166097; OTTHUMP00000166098; Protein Ash; SH2/SH3 adapter GRB2

Gene Aliases: AA408164; ASH; EGFRBP-GRB2; GRB2; Grb3-3; MST084; MSTP084; NCKAP2

UniProt ID: (Human) P62993, (Mouse) Q60631, (Rat) P62994

Entrez Gene ID: (Human) 2885, (Bovine) 535298, (Chinese hamster) 100769627, (Mouse) 14784, (Rat) 81504

Molecular Function: scaffold/adaptor protein