PA1-9051

antibody from Invitrogen Antibodies

Targeting: GRB2 NCKAP2

Antibody data

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Validation data

Product number: PA1-9051 - Provider product page
Provider: Invitrogen Antibodies
Product name: GRB2 Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Synthetic peptide
Description: This antibody is expected to recognize both reported isoforms (NP_002077.1 and NP_987102.1).
Concentration: 0.5 mg/mL

GRB2 protein structure - PA1-9051 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: 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) and A549 (Lane 9). The blots were probed with Goat Anti-GRB2 Polyclonal Antibody (Product # PA1-9051, 0.5 µg/mL) and detected by chemiluminescence using Rabbit anti-Goat IgG (H+L) Secondary Antibody, HRP conjugate (Product # 61-1620, 0.3 µg/mL, 1:5000 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).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of GRB2 was achieved by transfecting MCF7 cells with GRB2 specific siRNAs (Silencer® select Product # S6120 and s226232). Western blot analysis (Fig. a) was performed using membrane extracts from the GRB2 knock down cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with Anti-GRB2 Goat Polyclonal Antibody (Product # PA1-9051, 1 µg/mL) and Rabbit anti-Goat IgG (H+L) Superclonal™ Secondary Antibody, HRP (Product # A27014, 0.25 µg/mL, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Loss of signal upon siRNA mediated knock down confirms that antibody is specific to GRB2.