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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [2]
- Immunocytochemistry [2]
- Chromatin Immunoprecipitation [1]
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- Product number
- 701698 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- FOXA2 Recombinant Rabbit Monoclonal Antibody (9H5L7)
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- This antibody is predicted to react with Monkey, Horse, Cat and Dog.
- Antibody clone number
- 9H5L7
- Concentration
- 0.5 mg/mL
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on whole cell extracts (30 µg lysate) of Hep G2 (Lane 1), PC-3 (Lane 2) and COLO 205 (Lane 3). The blots were probed with Anti-FOXA2 Recombinant Rabbit Monoclonal Antibody (Product # 701698, 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 FOXA2 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® 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
- Western blot analysis was performed on whole cell extracts (30 µg lysate) of Hep G2 (Lane 1), PC-3 (Lane 2) and COLO 205 (Lane 3). The blots were probed with Anti-FOXA2 Recombinant Rabbit Monoclonal Antibody (Product # 701698, 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 FOXA2 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® 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
- Immunofluorescence was performed on methanol fixed HepG2 cells for detection of FOXA2 using FOXA2 Recombinant Rabbit Monoclonal Antibody (Product # 701698, 2 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Cytoskeleton was stained with alpha-Tubulin Monoclonal Antibody (Product # 32-2500, 1 µg/mL) followed by Goat anti-Mouse IgG Secondary Antibody, Alexa Fluor® 594 conjugate (Product # A-11032, 1:400). Panel a) shows representative cells that were stained for detection and localization of FOXA2 protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Panel c) represents cytoskeletal alpha-Tubulin staining (red). Panel d) is a composite image of Panels a, b and c clearly demonstrating nuclear localization of FOXA2. Panel e) represents control cells with no primary antibody to assess background.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- For immunofluorescence analysis, iPSC differentiated to Definitive Endoderm were fixed and permeabilized for detection of endogenous FOXA2 using anti-FOXA2 Recombinant Rabbit monoclonal Antibody (Product # 701698, 1:100 dilution) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2,000). Panel a) shows undifferentiated iPSC cells, Panel b) is the image of iPSC differentiated to Definitive Endoderm showing nuclear localization of FOXA2.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Enrichment of endogenous FOXA2 protein using Anti-FOXA2 Recombinant Rabbit Monoclonal Antibody: Chromatin Immunoprecipitation (ChIP) was performed using Anti-FOXA2 Recombinant Rabbit Monoclonal Antibody (Product # 701698, 5 µg) on sheared chromatin from 2 million HepG2 cells using the "MAGnify ChIP system" kit (Product # 49-2024). Normal Rabbit IgG (1 µg) was used as a negative IP control. The purified DNA was analyzed by 7500 Fast qPCR system (Product # 4351106) with optimized PCR primer pairs for the promoters of the active HNF1A region used as positive control target gene, and the region of the inactive SAT2 satellite repeat, used as negative control target gene. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.
