- MA5-36159 - Invitrogen Antibodies EIF3B antibody

Wu Z, Lei K, Xu S, He J, Shi E
Frontiers in immunology 2022;13:824946

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of eIF3b using a eIF3b monoclonal antibody (Product #MA5-36159). Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody at a dilution of 1:500 was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:5,000 dilution was used for 1 hour at room temperature. Positive control: Lane 1: A431 cell lysate, Lane 2: 293T cell lysate

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunocytochemical analysis of eIF3b in MCF-7 cells using a eIF3b monoclonal antibody (Product #MA5-36159). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1:1,000 dilution. The nuclear counter stain is DAPI.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunocytochemical analysis of eIF3b in MG-63 cells using a eIF3b monoclonal antibody (Product #MA5-36159). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1:1,000 dilution. The nuclear counter stain is DAPI.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemical analysis of eIF3b in paraffin-embedded human colon tissue using a monoclonal antibody (Product #MA5-36159). The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (1:50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemical analysis of eIF3b in paraffin-embedded human kidney tissue using a monoclonal antibody (Product #MA5-36159). The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (1:50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemical analysis of eIF3b in paraffin-embedded mouse testis tissue using a monoclonal antibody (Product #MA5-36159). The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (1:50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemical analysis of eIF3b in paraffin-embedded rat cerebellum tissue using a monoclonal antibody (Product #MA5-36159). The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (1:200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometric analysis of eIF3b using MCF-7 cells and a eIF3b monoclonal antibody (Product #MA5-36159). The cells were fixed, permeabilized and stained with the primary antibody at a dilution of 1:50 (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1:1000 dilution for 30 minutes. Unlabeled sample was used as a control (cells without incubation with primary antibody; black).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 6 EIF3B is a poor prognostic marker in melanoma. (A) Venn diagram shows the intersection of Cluster3 specific upregulated genes and the genes which are essential to melanoma human cell line growth using the DepMap database. (B) Kaplan-Meier analysis showing the association between the expression of EIF3B and SKCM patient overall survival (OS) in TCGA cohort (B) and GEO cohorts (C) , the Log-rank test was used to access the statistical differences. The differences of EIF3B expression between different subtypes in TCGA dataset (D) and GEO datasets (E) , the Anova test was used to calculate the statistical differences. (F) RT-qPCR and Western blot were used to verify the efficiency of EIF3B knockdown in SK-MEL-28 and A375 cell line. '*' indicates p-value

MA5-36159