- PA5-31177 - Invitrogen Antibodies EIF2S3 antibody

Submitted references Eif2s3y Promotes the Proliferation of Spermatogonial Stem Cells by Activating ERK Signaling.

Zhang M, Li N, Liu W, Du X, Wei Y, Yang D, Zhou Z, Ma F, Peng S, Zhang S, He X, Bai C, Li G, Hua J
Stem cells international 2021;2021:6668658

Suppression of MEHMO Syndrome Mutation in eIF2 by Small Molecule ISRIB.

Young-Baird SK, Lourenço MB, Elder MK, Klann E, Liebau S, Dever TE
Molecular cell 2020 Feb 20;77(4):875-886.e7

EIF2S3 Mutations Associated with Severe X-Linked Intellectual Disability Syndrome MEHMO.

Skopkova M, Hennig F, Shin BS, Turner CE, Stanikova D, Brennerova K, Stanik J, Fischer U, Henden L, Müller U, Steinberger D, Leshinsky-Silver E, Bottani A, Kurdiova T, Ukropec J, Nyitrayova O, Kolnikova M, Klimes I, Borck G, Bahlo M, Haas SA, Kim JR, Lotspeich-Cole LE, Gasperikova D, Dever TE, Kalscheuer VM
Human mutation 2017 Apr;38(4):409-425

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using eIF2 gamma Polyclonal Antibody (Product # PA5-31177). Sample (30 µg of whole cell lysate). Lane A: HepG2. 10% SDS PAGE. eIF2 gamma Polyclonal Antibody (Product # PA5-31177) diluted at 1:1,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using eIF2 gamma Polyclonal Antibody (Product # PA5-31177). Sample (30 µg of whole cell lysate). Lane A: HepG2. 10% SDS PAGE. eIF2 gamma Polyclonal Antibody (Product # PA5-31177) diluted at 1:1,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 1 The expression pattern of Eif2s3y in dairy goats. (a) Real-time PCR analysis of Eif2s3y expression levels in different tissues of adult dairy goats. All results were compared with the testis. (b) Real-time PCR analysis of Eif2s3y expression levels in the testes of dairy goats of different ages. All results were compared with that of 1-month-old goats. (c) Immunofluorescence staining of eIF2 gamma in the testes of 3-month-old dairy goats counterstained with Hoechst 33342. The white arrows indicated typical SSCs. Scale bars, 200 mu m (up) and 100 mu m (down). (d) Immunofluorescence staining of eIF2 gamma in the testes of 24-month-old dairy goats counterstained with Hoechst 33342. The white arrows indicated typical SSCs, and the red arrows indicated typical sperms. Scale bars, 200 mu m (up) and 100 mu m (down). Data are presented as mean +- SD and are represented by three independent repetitions; * P < 0.05, ** P < 0.01, and *** P < 0.001.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 3 Overexpression of Eif2s3y promotes the proliferation of dairy goat SSCs. (a, b) The schematic of lentivirus plasmid pCDH-CMV-MCS-EF1-puro and pCDH-CMV- Eif2s3y -EF1-puro. (c) Typical images of SSCs transfected with Control-Vector (left) or oeEif2s3y -Vector (right). Scale bar, 200 mu m. (d) Proliferation curve of Control and oeEif2s3y SSCs and the results of population doubling time (PDT) determination. (e) EdU incorporation assay of Control (up) and oeEif2s3y (down) SSCs. Scale bar, 400 mu m. (f) The ratio of EdU-positive cells to total cells. The proportion of positive cells is positively correlated with the cell proliferation rate. (g) RT-PCR analysis of the expression levels of Eif2s3y , Pcna , Cyclin D, and Zbtb16 in dairy goat SSCs transfected with Control-Vector or oeEif2s3y -Vector. (h) Western blotting detected the protein expression of ZBTB16, eIF2 gamma , PCNA, and Cyclin D in Control and oeEif2s3y SSCs. GAPDH was used as a loading control. (i) Gray intensity analysis of WB results normalized to GAPDH in (h). Data are presented as mean +- SD and are represented by three independent repetitions; * P < 0.05, ** P < 0.01, and *** P < 0.001.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 4 Depletion of Eif2s3y resulted in proliferation abnormality in goat SSCs. (a, b) The schematic of lentivirus plasmid CD513B-U6-MCS and CD513B-U6- shEif2s3y . (c) Typical images of shControl (left) and shEif2s3y (right) SSCs. Scale bar, 200 mu m. (d) Proliferation curve of shControl and shEif2s3y SSCs and the results of population doubling time (PDT) determination. (e) EdU incorporation assay of shControl (up) and shEif2s3y (down) SSCs. Scale bar, 400 mu m. (f) The ratio of EdU-positive cells to total cells. Data are presented as mean +- SD and are represented by three independent repetitions. (g) RT-PCR analysis of the expression levels of Eif2s3y , Pcna , Cyclin D, and Zbtb16 in shControl and shEif2s3y SSCs in vitro . (h) Western blotting detected the protein expression of ZBTB16, eIF2 gamma , PCNA, and Cyclin D in shControl and shEif2s3y SSCs. GAPDH was used as a loading control. (i) Gray intensity analysis of WB results normalized to GAPDH in (h). Data are presented as mean +- SD and are represented by three independent repetitions; * P < 0.05, ** P < 0.01, and *** P < 0.001.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 7 Eif2s3y promotes SSC proliferation by activating the ERK signaling pathway. Control and oeEif2s3y SSCs were treated with 1 mu M ERK pathway inhibitor PD0325901 or 10 mu M ERK pathway activator TPA for 24 h, respectively. shControl and shEif2s3y SSCs were treated with 10 mu M ERK pathway activator TPA for 24 h. DMSO was used for the control group. (a) EdU incorporation assay of Control and oeEif2s3y SSCs added with ERK pathway inhibitor PD0325901 or DMSO. Scale bar, 400 mu m. (b) The percentage of EdU-positive cells to total cells. The proportion of positive cells is positively correlated with the cell proliferation rate. (c) EdU incorporation assay of shControl and shEif2s3y SSCs added with ERK pathway activator TPA or DMSO. Scale bar, 400 mu m. (d) The percentage of EdU-positive cells to total cells. (e) Western blotting detected the protein expression of eIF2 gamma , PCNA, Cyclin D, pERK1/2, and ERK1/2 in SSCs treated as indicated. GAPDH was used as a loading control. (f) Gray intensity analysis of WB results normalized to GAPDH in (e). (g) Western blotting detected the protein expression of eIF2 gamma , PCNA, Cyclin D, pERK1/2, and ERK1/2 in SSCs treated as indicated. (h) Gray intensity analysis of WB results normalized to GAPDH in (g). Data are presented as mean +- SD and are represented by three independent repetitions; * P < 0.05, ** P < 0.01, and *** P < 0.001; N.S. means P >= 0.05.

PA5-31177

Antibody data