711581

antibody from Invitrogen Antibodies

Targeting: EIF2AK1 HRI, KIAA1369

Western blot (Supportive data in Antibodypedia)

Immunoprecipitation (Data presented on provider website)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

711581

Provider

Invitrogen Antibodies

Product name

HRI Recombinant Polyclonal Antibody (7HCLC)

Antibody type

Polyclonal

Antigen

Other

Reactivity

Human

Host

Rabbit

Isotype

IgG

Antibody clone number

7HCLC

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.

Western blot

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).

Other assay

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.

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.