Antibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [2]
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Validation data
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- Product number
- 720278 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- GHSR Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- These Polyclonal antibodies are of rabbit origin developed by immunizing animals with proteins or peptides. The polyclonal antibody is purified by affinity purification from the rabbit sera generated after immunizing the rabbits with a specific type of protein or peptide. The purified antibody is tested for its functionality in various relevant research applications. The antibody is developed for Research Use Only and is non-hazardous or non-infectious in nature.
- Concentration
- 0.5 mg/mL
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Supportive validation
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- Invitrogen Antibodies (provider)
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- Experimental details
- Western blot analysis was performed on tissue and membrane enriched cell extracts (30 µg lysate) of A549 (Lane 1), U-87 MG (Lane 2), Hep G2 (Lane 3), Mouse liver (Lane 4), Rat liver (Lane 5), SH-SY5Y (Lane 6), HEL 92.1.7 (Lane 7) and NTERA-2 (Lane 8). The blots were probed with Growth hormone secretagogue receptor 1a/GHSR1a Rabbit Polyclonal antibody (Product # 720278, 1-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A ~48 kDa band corresponding to Growth hormone secretagogue receptor 1a/GHSR1a was observed. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10% Bis-Tris gel (Product # NP0301BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with 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)
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- Experimental details
- Knockdown of GHSR was achieved by transfecting A549 cells with GHSR specific siRNAs (Silencer® select Product # s194432, s194431). Western blot analysis (Fig. a) was performed using whole cell extracts from the GHSR knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with GHSR Polyclonal Antibody (Product # 720278, 1 µg/ml) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25µg/ml, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to GHSR.