- AHO1182 - Invitrogen Antibodies EIF2S1 antibody

Submitted references Co-opting regulation bypass repair as a gene-correction strategy for monogenic diseases.

Hu J, Bourne RA, McGrath BC, Lin A, Pei Z, Cavener DR
Molecular therapy : the journal of the American Society of Gene Therapy 2021 Nov 3;29(11):3274-3292

Transmembrane Prolyl 4-Hydroxylase is a Novel Regulator of Calcium Signaling in Astrocytes.

Byts N, Sharma S, Laurila J, Paudel P, Miinalainen I, Ronkainen VP, Hinttala R, Törnquist K, Koivunen P, Myllyharju J
eNeuro 2021 Jan-Feb;8(1)

Maytansine-bearing antibody-drug conjugates induce in vitro hallmarks of immunogenic cell death selectively in antigen-positive target cells.

Bauzon M, Drake PM, Barfield RM, Cornali BM, Rupniewski I, Rabuka D
Oncoimmunology 2019;8(4):e1565859

Intermittent hypoxia leads to functional reorganization of mitochondria and affects cellular bioenergetics in marine molluscs.

Ivanina AV, Nesmelova I, Leamy L, Sokolov EP, Sokolova IM
The Journal of experimental biology 2016 Jun 1;219(Pt 11):1659-74

Long-Term Acclimation to Different Thermal Regimes Affects Molecular Responses to Heat Stress in a Freshwater Clam Corbicula Fluminea.

Falfushynska HI, Phan T, Sokolova IM
Scientific reports 2016 Dec 20;6:39476

Measurement of the unfolded protein response to investigate its role in adipogenesis and obesity.

Han J, Kaufman RJ
Methods in enzymology 2014;538:135-50

ER-stress-induced transcriptional regulation increases protein synthesis leading to cell death.

Han J, Back SH, Hur J, Lin YH, Gildersleeve R, Shan J, Yuan CL, Krokowski D, Wang S, Hatzoglou M, Kilberg MS, Sartor MA, Kaufman RJ
Nature cell biology 2013 May;15(5):481-90

Persistent redistribution of poly-adenylated mRNAs correlates with translation arrest and cell death following global brain ischemia and reperfusion.

Jamison JT, Kayali F, Rudolph J, Marshall M, Kimball SR, DeGracia DJ
Neuroscience 2008 Jun 23;154(2):504-20

Impaired dexamethasone-mediated induction of tryptophan 2,3-dioxygenase in heme-deficient rat hepatocytes: translational control by a hepatic eIF2alpha kinase, the heme-regulated inhibitor.

Liao M, Pabarcus MK, Wang Y, Hefner C, Maltby DA, Medzihradszky KF, Salas-Castillo SP, Yan J, Maher JJ, Correia MA
The Journal of pharmacology and experimental therapeutics 2007 Dec;323(3):979-89

No comments: Submit comment

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Proteins from cell extracts of human HeLa cells (lane 1), mouse 3T3L1 cells (lane 2), human Jurkat (lane 3), and rat L6 cells (lane 4) were resolved by SDS-PAGE and transferred to PVDF. The membranes were incubated with this antibody at a concentration of 1 µg/mL. After washing, the membranes were incubated with a goat F (ab')2 anti-rabbit IgG alkaline phosphatase conjugated secondary antibody (Product # ALI4405) at a 1:2000 dilution. Bands were detected with CDP-substrate using the WesternStar™ method (Tropix) and Kodak BioMax film.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on whole cell extracts (20 µg lysate) of COS-7 (Lane 1), MCF7 (Lane 2), PC-12 (Lane 3), HeLa (lane 4), Jurkat (lane 5), NIH/3T3 (lane 6), A-431 (lane 7), NTERA-2 (lane 8), SH-SY5Y (lane 9), KARPAS-299 (lane 10) and Daudi (lane 11). The blots were probed with Anti-eIF2a Rabbit Polyclonal Antibody (Product # AHO1182, 1:500 dilution) 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 36 kDa band corresponding to eIF2a was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0342BOX), 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® 2 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: Knockdown of Eukaryotic translation initiation factor 2 subunit 1 was achieved by transfecting HeLa with Eukaryotic translation initiation factor 2 subunit 1 specific siRNAs (Silencer® select Product # S4556, S4555). Western Blot analysis (Fig. a) was performed using Whole cell extracts from the Eukaryotic translation initiation factor 2 subunit 1 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The Blot was probed with eIF2a Polyclonal Antibody (Product # AHO1182, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution). Densitometric analysis of this western Blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to Eukaryotic translation initiation factor 2 subunit 1.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot was performed using Anti-eIF2a Polyclonal Antibody (Product # AHO1182) and a 38 kDa band corresponding to Eukaryotic translation initiation factor 2 subunit 1 was observed across tested cell lines with an uncharacterised band (*) at ~30 kDa. Whole cell extracts (40 µg lysate) of U-87 MG (Lane 1), HeLa (Lane 2), A-431 (Lane 3), HT-29 (Lane 4), MCF7 (Lane 5), Jurkat (Lane 6), NIH/3T3 (Lane 7), PC-12 (Lane 8) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # LC2001) by iBlot® 2 Dry Blotting System (Product # IB21001). The Blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of EIF2-alpha was done on 70% confluent log phase A549 cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with EIF2-alpha Rabbit Polyclonal Antibody (Product # AHO1182) at 1:250 dilution in1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing Cytoplasmic localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of EIF2-alpha was done on HeLa cells treated with Thapsigargin (50ng/mL, 20 minutes). Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Triton™ X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with EIF2-alpha Rabbit Polyclonal Antibody (AHO1182, red histogram) or with rabbit isotype control (pink histogram) at 3-5 ug/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor® 488 Goat Anti-Rabbit Secondary Antibody (A11008) at a dilution of 1:400 for 30 minutes at room temperature. The representative 10,000 cells were acquired and analyzed for each sample using an Attune® Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 5. Maytansine treatment promotes the phosphorylation of EIF2A in target cell lines. BT474 (a) and BJAB (b) cells in culture were either untreated (top) or treated with free maytansine (100 nM, bottom) for the times indicated. Cells were directly lysed into Laemmli buffer containing DTT, transferred to PVDF membranes, and probed by immunoblotting for EIF2A total protein (eIF2a) or Ser51 phosphorylated-EIF2a (P-eIF2A).

AHO1182

Antibody data