702551

antibody from Invitrogen Antibodies

Targeting: EIF2AK1 HRI, KIAA1369

Western blot (Supportive data in Antibodypedia)

Antibody Data

Product number

702551

Provider

Invitrogen Antibodies

Product name

HRI Recombinant Rabbit Monoclonal Antibody (7H3L3)

Antibody type

Monoclonal

Antigen

Other

Reactivity

Human

Host

Rabbit

Isotype

IgG

Antibody clone number

7H3L3

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.

References

Submitted references

ER stress-induced mediator C/EBP homologous protein thwarts effector T cell activity in tumors through T-bet repression.

Cao Y, Trillo-Tinoco J, Sierra RA, Anadon C, Dai W, Mohamed E, Cen L, Costich TL, Magliocco A, Marchion D, Klar R, Michel S, Jaschinski F, Reich RR, Mehrotra S, Cubillos-Ruiz JR, Munn DH, Conejo-Garcia JR, Rodriguez PC
Nature communications 2019 Mar 20;10(1):1280

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 uM 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 uM for 1 hr) (Lane 8). The blots were probed with Anti-eIF2AK1/Heme regulated inhibitor (HRI) Recombinant Rabbit Monoclonal Antibody (Product # 702551, 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 eIF2AK1/Heme regulated inhibitor (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).