- PA5-27366 - Invitrogen Antibodies EIF2S1 antibody

Molagoda IMN, Kavinda MHD, Choi YH, Lee H, Kang CH, Lee MH, Lee CM, Kim GY
Antioxidants (Basel, Switzerland) 2021 Sep 18;10(9)

Karunarathne WAHM, Molagoda IMN, Lee KT, Choi YH, Yu SM, Kang CH, Kim GY
Antioxidants (Basel, Switzerland) 2021 Apr 9;10(4)

Cheung CHY, Hsu CL, Lin TY, Chen WT, Wang YC, Huang HC, Juan HF
Journal of biomedical science 2020 Jun 23;27(1):75

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of EIF2 alpha using 30 µg of H1299 lysate. Samples were loaded onto a 12% SDS-PAGE gel and probed with an EIF2 alpha polyclonal antibody (Product # PA5-27366) at a dilution of 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of eIF2a was performed by separating 30 µg of various whole cell extracts by 10% SDS-PAGE. Proteins were transferred to a membrane and probed with a eIF2a Polyclonal Antibody (Product # PA5-27366) at a dilution of 1:5000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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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 # PA5-27366, 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 # PA5-27366) and a 38 kDa band corresponding to Eukaryotic translation initiation factor 2 subunit 1 was observed across tested cell lines. 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) 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 Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunocytochemistry-Immunofluorescence analysis of eIF2a was performed in HCT 116 cells fixed in 4% paraformaldehyde at RT for 15 min. Green: eIF2a Polyclonal Antibody (Product # PA5-27366) diluted at 1:500. Blue: Hoechst 33342 staining.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunocytochemistry-Immunofluorescence analysis of eIF2a was performed in HeLa cells fixed in 4% paraformaldehyde at RT for 15 min. Green: eIF2a Polyclonal Antibody (Product # PA5-27366) diluted at 1:5000. Blue: Hoechst 33342 staining. Scale bar = 10 µm.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 8 Effect of anthocyanins from the flower petals of Hibiscus syriacus L. (Malvaceae, AHs) on ER stress and mtROS production. ( A ) and ( B ) The expression of GRP78, ATF4, p-eIF1alpha, CHOP, and beta-actin protein (left) and relative density (right). ( C ) The staining of MitoSOX Red and MitoTracker Green. ( D ) Survival rate of zebrafish. * p < 0.05 and *** p < 0.001 vs. UVB-irradiated cells and ### p < 0.001 vs. untreated cells (UT).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 4 Fisetin inhibits PM 2.5 -induced apoptosis by alleviating ER stress. HaCaT keratinocytes were treated with fisetin (0-10 uM) for 2 h and subsequently exposed to 100 ug/mL PM 2.5 for 24 h. ( A ) The total proteins were extracted, and Western blotting was performed for detecting the expression of GRP78, p-eIF2alpha, eIF2alpha, ATF4, and CHOP. beta-Actin was used as the loading control. ( B ) The cells were treated with 10 uM fisetin or 20 uM salubrinal in the presence or absence of 100 ug/mL PM 2.5 for 24 h. The cells were incubated with Ca 2+ -sensitive Fluo-4 AM for 10 min and live images were captured using a CELENA S Digital Imaging System. Scale bar = 100 um. ( C , D ) In a parallel experiment, the cells were treated with 10 uM fisetin or 20 uM salubrinal in the presence or absence of 100 ug/mL PM 2.5 for 24 h. The cells were stained with a ( C ) Muse Oxidative Stress Assay Kit and ( D ) Muse Annexin V & Dead Cell Assay Kit. *** p < 0.001 vs. untreated cells and # p < 0.05 vs. PM 2.5 -treated cells.

PA5-27366