Eif2a

Manufactured by Santa Cruz Biotechnology

Sourced in China, United States

EIF2a is a protein involved in the initiation of protein synthesis. It plays a crucial role in the regulation of translation by controlling the binding of messenger RNA to the ribosome. EIF2a is a key component of the eukaryotic translation initiation complex.

Automatically generated - may contain errors

Lab products found in correlation

Grp78, by Santa Cruz Biotechnology (2 mentions) P erk, by Santa Cruz Biotechnology (2 mentions) Dexamethasone (dex), by Selleck Chemicals (1 mentions) Cleaved caspase 3, by Cell Signaling Technology (1 mentions) P38 mapk, by Cell Signaling Technology (1 mentions) Psmd3, by Santa Cruz Biotechnology (1 mentions) Psmc5, by Santa Cruz Biotechnology (1 mentions) P p38mapk t180 y182, by Cell Signaling Technology (1 mentions) P eif2α s51, by Cell Signaling Technology (1 mentions) Xbp1s, by Cell Signaling Technology (1 mentions) Gapdh, by Proteintech (1 mentions) Edta free protease inhibitor, by Roche (1 mentions) Puromycin, by Merck Group (1 mentions) 3 methyladenine, by Merck Group (1 mentions) Hoechst pi, by Beyotime (1 mentions) Reactive oxygen species assay kit, by Beyotime (1 mentions) Hydroxychloroquine, by Tokyo Chemical Industry (1 mentions) N acetyl l cysteine, by Beyotime (1 mentions) Bcl2 associated x (bax), by Cell Signaling Technology (1 mentions) Caspase 8, by Cell Signaling Technology (1 mentions)

Spelling variants of this product

P-eIF2a (2 citations) Anti-eIF2a (2 citations)

7 protocols using eif2a

Comprehensive Proteostasis Profiling Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

BTZ (catalog S1013), CFZ (catalog S2853), MEL (catalog S8266), DEX (catalog S1322), and DOX (catalog S1208) were purchased from Selleck Chemicals. Recombinant human PSMB5 protein was custom-made by MedChemExpress. Recombinant human ISG20L2 protein was custom-made by Merry Bio Technology Co., Ltd.
Western blot antibodies against PSMB5 (catalog sc-393931), PSMD8 (catalog sc-514053), PSMD3 (catalog sc-393588), PSMC5 (catalog sc-390631), and eIF2a (catalog sc-133132) were purchased from Santa Cruz Biotechnology. Western blot antibodies against ISG20L2 (catalog 24639-1-AP) and GAPDH (catalog 60004-1-Ig) were purchased from Proteintech. Antibodies against ATF4 (catalog 11815), ATF6 (catalog 65880), cleaved caspase-3 (catalog 9661), p-p38MAPK (T180/Y182) (catalog 9211), p38MAPK (catalog 9212), p-eIF2α (S51) (catalog 3398), and XBP-1s (catalog 27901) were obtained from Cell Signaling Technology. Western blot antibodies against p-PERK (T982) (catalog WL05295), PERK (catalog WL03378), and CHOP (catalog WL00880) were obtained from Wanleibio.

Yang Y., Gao Y., Huang J., Yang Z., Luo H., Wang F., Xu J., Cui Y., Ding H., Lin Z., Zhai X., Qu Y., Zhang L., Liu T., Ye L., Niu T, & Zheng Y. (2022). ISG20L2 suppresses bortezomib antimyeloma activity by attenuating bortezomib binding to PSMB5. JCI Insight, 7(19), e157081.

+ Open protocol

+ Expand

Measuring Global Translation Levels

Check if the same lab product or an alternative is used in the 5 most similar protocols

U2OS cells (WT or G3BP1/2 KO) were grown and treated as described previously. Cells were stressed with 1 mM sodium arsenate and allowed to recover in fresh media for the indicated times. Before collection, cells were treated with 91 μM of Puromycin for 5 min at 37°C. As control for specific Puromycin incorporation into nascent polypeptide chains, cells were pre-treated with 2 μg/ml Harringtonine for 15 min before Puromycin addition. After treatment, cells were washed with PBS and lysed with RIPA buffer (25 mM Tris/HCl pH 7.5, 150 mM NaCl, 1% NP-40, 0.1% SDS, EDTA-free protease inhibitors (Roche Applied Sciences)). Cell extracts were centrifuged 5 min at maximum speed and 4°C and the supernatant was recovered. 10 μg of protein extracts were loaded into SDS-PAGE and analyzed by immunoblot. Antibodies against Puromycin (Merck Millipore), p(S52)-eIF2a (Thermo Fisher Scientific) and eIF2a (Santa Cruz Biotechnology) were used to assess translation levels and follow stress kinetics. Relative translation levels (Puromycin signal) were quantified using Fiji and Excel.

Guillén-Boixet J., Kopach A., Holehouse A.S., Wittmann S., Jahnel M., Schlüßler R., Kim K., Trussina I.R., Wang J., Mateju D., Poser I., Maharana S., Ruer-Gruß M., Richter D., Zhang X., Chang Y.T., Guck J., Honigmann A., Mahamid J., Hyman A.A., Pappu R.V., Alberti S, & Franzmann T.M. (2020). RNA-Induced Conformational Switching and Clustering of G3BP Drive Stress Granule Assembly by Condensation. Cell, 181(2), 346-361.e17.

+ Open protocol

+ Expand

Investigating Cellular Stress Responses

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cell culture media and additives were obtained from the HyClone Company (Beijing, China). N-acetyl-L-cysteine (>99%), Salubrinal, Hoechst/PI and a Reactive Oxygen Species Assay Kit were purchased from Beyotime Biotechnology (Shanghai, China). The 3-methyladenine was purchased from Sigma-Aldrich (Shanghai, China). Hydroxychloroquine was purchased from Tokyo Chemical Industry (Shanghai, China). GSK2606414 (>99.38) was purchased from Med Chen Express (New Zealand, USA). PERK (Cat. No. 3192), IRE1a (Cat. No. 3294) and p62(Cat. No. 5114) antibodies were purchased from Cell Signaling Technology (Shanghai, China). EIF2a (Cat. No.sc-133132) antibodies were acquired from Santa Cruz Biotechnology (Beijing, China). Rhodopsin (Cat. No. OM186133) antibodies were purchased from OmnimAbs (Shanghai, China). LC3B (Cat. No. ab192890) antibodies were purchased from Abcam (Cambridge, MA, USA). BECN1 (Cat. No. 11306-1-AP) antibodies were purchased from Proteintech (Wuhan, China). P-IRE1a (Cat. No. 13013), p-PERK (Cat. No. 12814), ATF6 (Cat. No. 24382), p-EIF2a (Cat. No. 11279), ATF4 (Cat. No. 32007), CHOP (Cat. No. 40744), RPE65(Cat. No. 49495), β-actin (Cat. No. 21800) and secondary antibodies were obtained from Signalway Technology (St. Louis, MO, USA).

Song J.Y., Fan B., Che L., Pan Y.R., Zhang S.M., Wang Y., Bunik V, & Li G.Y. (2020). Suppressing endoplasmic reticulum stress-related autophagy attenuates retinal light injury. Aging (Albany NY), 12(16), 16579-16596.

+ Open protocol

+ Expand

Profiling Apoptosis Signaling Pathways

Check if the same lab product or an alternative is used in the 5 most similar protocols

Western blot was performed on purified 5T33MM cells as described previously [12 ] using the following antibodies: pJNK (Thr183/Tyr185), JNK, MCL-1, Bim (EL/L/S), Bax, Caspase-8, Caspase-9, Caspase-3, PARP, ATF4, Caspase-12, XBP1, CHOP and β-tubulin, all purchased from Cell Signaling Technology (Boston, MA, USA). PERK, IRE1, GRP-78, pEIF2A (Ser52) and EIF2A were purchased from Santa Cruz Biotechnology Inc. (Dallas, TX, USA) and Ubiquitin from Enzo Life Sciences (Farmingdale, NY, USA).

De Beule N., Menu E., Bertrand M.J., Favreau M., De Bruyne E., Maes K., De Veirman K., Radwanska M., Samali A., Magez S., Vanderkerken K, & De Trez C. (2017). Experimental African trypanosome infection suppresses the development of multiple myeloma in mice by inducing intrinsic apoptosis of malignant plasma cells. Oncotarget, 8(32), 52016-52025.

+ Open protocol

+ Expand

Western Blot Analysis of ER Stress Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols

Heart samples were homogenized in RIPA buffer containing 50-mM Tris (pH 7.4), 1% Triton C-100, and 150-mM NaCl, and freshly added phenylmethylsulfonyl fluoride (PMSF; 0.1%) protease inhibitor (Sigma-Aldrich). Final supernatants were obtained by centrifugation at 12,000 × g for 10 min at 4°C and protein concentrations were determined using a standard BCA assay kit. Proteins (40 μg) were then electrophoresed on 10%-12% sodium dodecyl sulfate polyacrylamide gels and were transferred to polyvinylidene difluoride (PVDF) membranes (Millipore). Membranes were then blocked in Tris-buffered saline containing Tween-20 (TBST; 20-mM Tris, 137-mM NaCl, and 0.1% Tween-20) and 5% non-fat milk at room temperature for 1 h. PVDF membranes were then incubated with primary antibodies against GRP78 (1:500, Santa Cruz), PERK (1:1000, Santa Cruz), P-PERK (1:1000, Santa Cruz), eIF2a (1:1000, Santa Cruz), P-eIF2a (1:1000, Santa Cruz), CHOP (1:500; Abcam), Caspase-12 (1:500, Santa Cruz) and NAPDH (1:4000, Santa Cruz) overnight at 4°C. After washing in TBST, membranes were subjected to appropriate secondary antibodies for 1 h at 37°C, and bands from heart samples were visualized using enhanced chemiluminescence (ECL) detection kits and were quantified using image J software.

Bi F., Xu Y., Chen G, & Wang P. (2020). Anti-inflammatory and Anti-endoplasmic reticulum stress Effects of catalpol Against myocardial ischemia-reperfusion injury in streptozotocin-induced diabetic rats. Anais da Academia Brasileira de Ciencias, 92(4).

+ Open protocol

+ Expand

Immunoblot Analysis of eIF2α Signaling

Check if the same lab product or an alternative is used in the 5 most similar protocols

For immunoblot analysis, HeLa cell cultures were placed on ice and lysed in ice-cold 1% CHAPSO, 150 mM NaCl, 20 mM K-HEPES, pH 7.5, 10% glycerol, 1 mM EDTA, 100 mM NaF, 17.5 mM B-glycerophosphate, and 1X protease inhibitor cocktail for 5 min. Lysates were TCA precipitated as noted above, washed with 100% acetone, resuspended in 0.5 M Tris, pH 11, 5% SDS, and protein concentrations determined via BCA assay (Pierce). Equivalent protein mass per sample was separated by SDS-PAGE, transferred to nitrocellulose membranes, blocked in 10% nonfat dry milk/Tris-buffered saline and processed per antibody supplier's recommendations.
Antibodies used include eIF2a (Santa Cruz, Cat. No. sc11386; 1:500 dilution) and phospho-eIF2a (Cell Signaling Technologies, Cat. No. 3398; 1:1500 dilution).

Child J.R., Chen Q., Reid D., Jagannathan S., & Nicchitta C.V. (2021). Selective recruitment of endoplasmic reticulum-targeted and cytosolic mRNAs into membrane-associated stress granules.

+ Open protocol

+ Expand

Modulation of TNF-β Signaling by eIF2α

Check if the same lab product or an alternative is used in the 5 most similar protocols

BHK-21 cell cultures were transiently transfected with >75% efficiency using Turbofect (Fermentas), with 2.5 mg wild-type (WT) or mutant TNF-b/2-APRE or TNF-b DNA or with 1.5 mg each of TNF-b/2-APRE or TNF-b DNA and eIF2aWT, eIF2aS51A, or eIF2aS51D expression vector, or pBS. In all transfections, 0.5 mg pEGFP-N3 DNA (Clontech Laboratories) was included to assess transfection efficiency. TNF-b and TNF-a protein secreted into culture medium was quantitated by ELISA (DuoSet, R&D Systems). Lysates from cells treated with salubrinal (Calbiochem) or DMSO solvent were immunoblotted with antibodies for P-eIF2a (Cell Signaling Technology) and eIF2a (Santa Cruz Biotechnology).

Namer L.S., Osman F., Banai Y., Masquida B., Jung R, & Kaempfer R. (2017). An Ancient Pseudoknot in TNF-α Pre-mRNA Activates PKR, Inducing eIF2α Phosphorylation that Potently Enhances Splicing. Cell reports, 20(1).

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!