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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [2]
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- Product number
- 702742 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- HK1 Recombinant Rabbit Monoclonal Antibody (18H16L7)
- Antibody type
- Monoclonal
- Antigen
- Synthetic peptide
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Antibody clone number
- 18H16L7
- Vial size
- 100 µg
- Concentration
- 0.5 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of Hexokinase 1 was achieved by transfecting A549 cells with Hexokinase 1 specific validated siRNA (Silencer® select Product # s6558). Western blot analysis (Fig a) was performed using whole cell extracts (1% SDS) from Hexokinase 1 knockdown cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blots were probed with Anti-Hexokinase 1 Recombinant Rabbit Monoclonal Antibody (Product # 702742, 1-3 µg/mL) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 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 Hexokinase 1.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on Whole cell extracts (60 µg lysate) of PC-3 (Lane 1), K-562 (Lane 2), U-87 MG (Lane 3), MCF-7 (Lane 4), A-431 (Lane 5) PANC-1 (Lane 6) and A549 (Lane 7). The blots were probed with Anti-Hexokinase 1 Recombinant Rabbit Monoclonal Antibody (Product # 702742, 2.5 µg/mL) 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 102 kDa band corresponding to Hexokinase 1 was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex®NuPAGE®4-12% Bis-Tris gel (Product # NP0322BOX), 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® 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).
