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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [1]
- Other assay [2]
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Validation data
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- Product number
- 711581 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- HRI Recombinant Polyclonal Antibody (7HCLC)
- Antibody type
- Polyclonal
- Antigen
- Other
- Description
- This antibody is predicted to react with Monkey, Horse, Mouse
- Antibody clone number
- 7HCLC
- 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 Membrane extracts (30 µg lysate) of K-562 (Lane 1), K-562 treated with H2O2 (150 µM for 1 hr) (Lane 2), HEL 92.1.7 (Lane 3), HEL 92.1.7 treated with H2O2 (150 uM for 1 hr) (Lane 4), T-47D (Lane 5), T-47D treated with H2O2 (150 uM for 1 hr) (Lane 6), HEK-293 (Lane 7) and HEK-293 treated with H2O2 (150 µM for 1 hr) (Lane 8). The blots were probed with Anti-HRI Recombinant Rabbit Polyclonal Antibody (Product # 711581, 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 71 kDa band corresponding to HRI was observed across the cell lines (with treatment difference) tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris gel (Product # NP0321BOX), 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
- Immunoprecipitation of HRI was performed in K562 cells. Antigen-antibody complexes were formed by incubating approximately 500 µg whole cell lysate with 5 to 10 µL of recombinant polyclonal HRI antibody (Product # 711581) rotating 60 min at RT. The immune complexes were captured on 625 µg of anti-rabbit coated Dynabeads (Product # 11204D) and washed extensively. They were then eluted and analyzed using the Simple Western system using the same antibody as used in immunoprecipitation at a dilution of 1:25, followed by a 1:100 dilution of secondary antibody. Lane 1 is the input, lane 2 no antibody IP and lane 3 is the target specific IP. Data courtesy of the Yeo lab as part of the ENCODE project.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunoprecipitation of HRI was performed in K562 cells. Antigen-antibody complexes were formed by incubating approximately 500 µg whole cell lysate with 5 to 10 µL of recombinant polyclonal HRI antibody (Product # 711581) rotating 60 min at RT. The immune complexes were captured on 625 µg of anti-rabbit coated Dynabeads (Product # 11204D) and washed extensively. They were then eluted and analyzed using the Simple Western system using the same antibody as used in immunoprecipitation at a dilution of 1:25, followed by a 1:100 dilution of secondary antibody. Lane 1 is the input, lane 2 no antibody IP and lane 3 is the target specific IP. Data courtesy of the Yeo lab as part of the ENCODE project.
