PA5-62621

antibody from Invitrogen Antibodies

Targeting: GSPT1 eRF3a, ETF3A, GST1

Western blot (Data presented on provider website)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Data presented on provider website)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-62621

Provider

Invitrogen Antibodies

Product name

GSPT1 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant full-length protein

Description

Immunogen sequence: GGAANNHGAG SGAGGRAAPV ESSQEEQSLC EGSNSAVS Highest antigen sequence identity to the following orthologs: Mouse - 66%, Rat - 74%.

Reactivity

Human

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

0.10 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

References

Submitted references

Effect of small molecule eRF3 degraders on premature termination codon readthrough.

Baradaran-Heravi A, Balgi AD, Hosseini-Farahabadi S, Choi K, Has C, Roberge M
Nucleic acids research 2021 Apr 19;49(7):3692-3708

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent staining of GSPT1 in human cell line HeLa shows positivity in cytoplasm & vesicles. Samples were probed using a GSPT1 Polyclonal Antibody (Product # PA5-62621).

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 1. Effect of siRNA knockdown of eRF3a and eRF3b on TP53 PTC readthrough. ( A ) HDQ-P1 cells with a homozygous nonsense mutation in the TP53 gene (R213X) were transfected with either non-target siRNA or several ON-TARGETplus siRNAs targeting eRF3a . Levels of SURF complex components eRF3a, eRF1, UPF1 and SMG1 were measured 96 h after transfection. GAPDH was used as loading control. ( B ) p53 levels (full-length, FL-p53; truncated, TR-p53) were measured in the same extracts as in A . Vinculin was used as loading control. ( C ) p53 and vinculin levels were measured in transfected cells exposed to 20 mug/ml G418 for 48 h. ( D ) eRF3a and eRF3b levels were measured in HDQ-P1 and H1299-p53 R213X cell extracts. ( E, F ) H1299-p53 R213X cells were transfected with non-target siRNA or several ON-TARGETplus siRNAs targeting eRF3a ( E ) or eRF3b ( F ). eRF3a and eRF3b levels were measured 96 h after transfection. 48 h after transfection, samples were exposed to 20 mug/ml G418 for another 48 h and p53 and vinculin levels were determined. Protein levels in all panels were measured using automated capillary electrophoresis western analysis. In this and subsequent figures, equal amounts of protein lysate were loaded in all capillaries in each panel, except where indicated. The amounts of eRF3a, eRF3b, eRF1, UPF1 and SMG1 are expressed as percentage of levels in untreated cells. As FL-p53 is essentially undetectable in untreated HDQ-P1 cells in most experiments, FL-p53 levels are expr