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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [5]
- Immunohistochemistry [4]
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- 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). This antibody is predicted to react with bovine and canine based on sequence homology. This antibody is tested in Peptide ELISA: antibody detection limit dilution 32,000.
- Reactivity
- Human, Mouse, Rat, Porcine
- Host
- Goat
- Isotype
- IgG
- Vial size
- 100 μg
- Concentration
- 0.5 mg/mL
- Storage
- -20°C, Avoid Freeze/Thaw Cycles
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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).
- 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 Heavy Chain 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.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of GRB2 using GRB2 Polyclonal Antibody (Product # PA1-9051) (0.05 µg/mL) in staining of Mouse (A+C) and Rat (B+D) Brain (A+B) and Spleen (C+D) lysates (35 µg total protein in RIPA buffer). Primary incubated for 1 hour. Detected by western blot using chemiluminescence.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of GRB2 using GRB2 Polyclonal Antibody (Product # PA1-9051) (0.03 µg/mL) in staining of Human (A) and Pig (B) Spleen lysates (35 µg protein in RIPA buffer). Primary incubation was 1 hour. Detected by chemiluminescence.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of GRB2 using GRB2 Polyclonal Antibody (Product # PA1-9051) (0.05 µg/mL) in staining of Human Thymus (A) and MOLT4 (B) lysates (35 µg total protein in RIPA buffer). Primary incubated for 1 hour. Detected by chemiluminescence.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of GRB2 in human pancreas. Samples were incubated with GRB2 polyclonal antibody (Product # PA1-9051) using a dilution of 5 µg/mL. Formalin-fixed, paraffin-embedded tissue after heat-induced antigen retrieval.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry (PFA fixed) analysis of GRB2 using GRB2 Polyclonal Antibody (Product # PA1-9051) (1 µg/mL) in staining of paraffin embedded Human Lymph Node. Microwaved antigen retrieval with citrate buffer pH 6, HRP-staining.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of GRB2 in human pancreas. Samples were incubated with GRB2 polyclonal antibody (Product # PA1-9051) using a dilution of 5 µg/mL. Formalin-fixed, paraffin-embedded tissue after heat-induced antigen retrieval.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry (PFA fixed) analysis of GRB2 using GRB2 Polyclonal Antibody (Product # PA1-9051) (1 µg/mL) in staining of paraffin embedded Human Lymph Node. Microwaved antigen retrieval with citrate buffer pH 6, HRP-staining.
