Anti-GRSF1 抗体 [EPR16679]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRSF1 antibody [EPR16679] (AB194358)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling GRSF1 with ab194358 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Counter stained with Hematoxylin.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-GRSF1 antibody [EPR16679] (AB194358)

Intracellular Flow Cytometry analysis of HeLa cells labelling GRSF1 (red) with purified ab194358 at dilution of 1/250. The secondary antibody used was Alexa Fluorr^® 488 goat-anti-rabbit IgG (1/2000). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Isotype control antibody was Rabbit monoclonal IgG (black). The blue line shows cells without incubation with primary antibody and secondary antibody.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-GRSF1 antibody [EPR16679] (AB194358)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized HeLa cells labeling GRSF1 with ab194358 at 1/100 dilution followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/400 dilution. The nuclear counter stain is DAPI (blue).

• WB

Supplier Data

Western blot - Anti-GRSF1 antibody [EPR16679] (AB194358)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-GRSF1 antibody [EPR16679] (ab194358) at 1/10000 dilution

Lane 1:

HeLa cell lysate at 20 µg

Lane 2:

293 cell lysate at 20 µg

Lane 3:

HepG2 cell lysate at 20 µg

Lane 4:

Jurkat cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 53 kDa

Observed band size: 53 kDa

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• WB

CiteAb

Western blot - Anti-GRSF1 antibody [EPR16679] (AB194358)

GRSF1 western blot using anti-GRSF1 antibody [EPR16679] ab194358. Publication image and figure legend from Varshney, D., Cuesta, S. M., et al., 2021, Sci Rep, PubMed 34815422.

ab194358 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab194358 please see the product overview.

RNA G4s in ribosomal protein mRNA regulate translation. (a) Example snapshots demonstrating BG4 peaks in 5' UTRs and overlapping iCLAE peaks. BG4 track depict logFC of BG4 IP vs input and iCLAE track depict counts per million. (b) Western blot analysis of ribosomal protein levels following shRNA mediated depletion of DDX3X, DHX36 and GRSF1. Actin serves as loading control. Number below each lane represents fold change in ribosomal protein levels compared to non-targeting control (NTC) when normalised to actin. (c) Fold change in luminescence comparing in vitro translation of luciferase gene from wild-type ribosomal protein UTRs containing G4 (UTRQ) to mutant UTRs with G4 disruption (MutQ). (d) UV absorption at 254 nM following sucrose fractionation for polysome profiling of cells treated with DMSO (black) and 2 μM PDS (blue) for 45 min. Monosomal and polysomal fractions are highlighted. (e) Fold change in translation efficiency (logFC TE) of each transcript plotted against the significance (- log10 p-value) over three independent biological replicates. All ribosomal protein mRNAs are highlighted in red and those with significant changes (FDR < 0.1) are labelled with the corresponding protein names.

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