HEK-293 treated with 2μg/mL Tunicamycin for 8 hours cell lysate, C6 treated with 2μg/mL Tunicamycin for 8 hours cell lysate, HeLa treated with 2μg/mL Tunicamycin for 8 hours cell lysate, HeLa treated with 250μM CoCl2 for 6 hours cell lysate, NIH/3T3 treated with 2μg/mL Tunicamycin for 8 hours cell lysate.
Conjugation
unconjugated
Clone Number
PSH07-08
Reactivity Data
Tested Verified (internally validated)
Published Reported in literature (not internally validated)
Predicted Predicted reactive (based on sequence homology)
Not recommended Not recommended (failed internal validation)
WB
IP
Human
Mouse
Rat
Product Features
Form
Liquid
Concentration
1 mg/mL.(The concentration of this product may be batch-dependent)
Lot Concentration Lookup
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Activating transcription factor 4 (tax-responsive enhancer element B67), also known as ATF4, is a protein that in humans is encoded by the ATF4 gene. ATF4 transcription factor is also known to play role in osteoblast differentiation along with RUNX2 and osterix. Terminal osteoblast differentiation, represented by matrix mineralization, is significantly inhibited by the inactivation of JNK. JNK inactivation downregulates expression of ATF-4 and, subsequently, matrix mineralization. IMPACT protein regulates ATF4 in C. elegans to promote lifespan. ATF4 is also involved in the cannabinoid Δ9-tetrahydrocannabinol–induced apoptosis in cancer cells, by the proapoptotic role of the stress protein p8 via its upregulation of the endoplasmic reticulum stress-related genes ATF4, CHOP, and TRB3.
Background References
1. He F et al. ATF4 suppresses hepatocarcinogenesis by inducing SLC7A11 (xCT) to block stress-related ferroptosis. J Hepatol. 2023 Aug
2. Xiao Y et al. Advances in the roles of ATF4 in osteoporosis. Biomed Pharmacother. 2023 Dec
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722796) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
☑ Cell treatment (CT)
Western blot analysis of ATF4 on different lysates with Rabbit anti-ATF4 antibody (HA722796) at 1/2,000 dilution.
Lane 1: HeLa cell lysate (20 µg/Lane) Lane 2: HeLa treated with 2μg/mL Tunicamycin for 8 hours cell lysate (20 µg/Lane) Lane 3: HeLa cell lysate (20 µg/Lane) Lane 4: HeLa treated with 250μM CoCl2 for 6 hours cell lysate (20 µg/Lane) Lane 5: NIH/3T3 cell lysate (20 µg/Lane) Lane 6: NIH/3T3 treated with 2μg/mL Tunicamycin for 8 hours cell lysate (20 µg/Lane)
Predicted band size: 39 kDa Observed band size: 50 kDa
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722796) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
☑ Cell treatment (CT)
ATF4 was immunoprecipitated from 0.2 mg HeLa treated with 2μg/mL Tunicamycin for 8 hours cell lysate with HA722796 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA722796 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: HeLa treated with 2μg/mL Tunicamycin for 8 hours cell lysate (input) Lane 2: HA722796 IP in HeLa treated with 2μg/mL Tunicamycin for 8 hours cell lysate Lane 3: Rabbit IgG instead of HA722796 in HeLa treated with 2μg/mL Tunicamycin for 8 hours cell lysate