Mouse anti g3bp1

Manufactured by Abcam

Mouse anti-G3BP1 is an antibody that specifically binds to the G3BP1 protein. G3BP1 is an RNA-binding protein involved in the formation of stress granules, which are cytoplasmic aggregations of stalled translation initiation complexes, mRNA, and RNA-binding proteins that assemble in response to cellular stresses. This antibody can be used to detect the G3BP1 protein in various research applications.

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Lab products found in correlation

Goat anti tia1, by Santa Cruz Biotechnology (2 mentions) Hoechst 33342, by Merck Group (2 mentions) Puromycin, by Thermo Fisher Scientific (1 mentions) Sodium arsenite, by Merck Group (1 mentions) Mouse anti gapdh, by Abcam (1 mentions) Pkr inhibitor c16, by Merck Group (1 mentions) Isrib, by Merck Group (1 mentions) Goat anti eif3η, by Santa Cruz Biotechnology (1 mentions) Rabbit anti actin, by Merck Group (1 mentions) Rabbit anti calnexin, by Abcam (1 mentions) Poly 1 c, by Merck Group (1 mentions) Mouse anti puromycin, by Kerafast (1 mentions) Rabbit anti pabp, by Abcam (1 mentions) Mowiol, by Merck Group (1 mentions) Mouse anti polyq, by Merck Group (1 mentions) Alexa fluor 568f ab 2 fragment of goat anti rabbit igg h l, by Thermo Fisher Scientific (1 mentions) Alexa fluor 594 goat anti rabbit igg h l, by Thermo Fisher Scientific (1 mentions) Alexa fluor 488 goat anti mouse, by Thermo Fisher Scientific (1 mentions) Hoechst 33342, by Thermo Fisher Scientific (1 mentions) Alexa488 labeled goat anti mouse igg, by Abcam (1 mentions)

4 protocols using mouse anti g3bp1

Molecular Insights into Viral Stress Response

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Sodium arsenite (Sigma-Aldrich) was added to the cells at a concentration of 250 μM for 20 min prior to fixation or cell lysate collection. The PKR inhibitor C16 (Sigma-Aldrich) was added to infected cells at a concentration of 1 μM at 1 hpi, and cell lysates were collected at 12 hpi. The ISR inhibitor ISRIB (Sigma-Aldrich) was added at 1 hpi at 0.5 μM. Puromycin (Life Technologies) was added to cells at a concentration of 10 μg/ml at indicated times prior to cell lysate collection. Poly(I⋅C) (Sigma-Aldrich, catalog no. P1530) is a dsRNA analogue and was used as a positive control for immune activation. Poly(I⋅C) was used at a concentration of 20 μg/ml and added to the cell culture medium in combination with Lipofectamine 2000. MNV ORF1 plasmids were described and used previously (22 (link)).
Goat anti-eIF3η, goat anti-G3BP1, and goat anti-TIA-1 were all purchased from Santa Cruz Biotech. Rabbit anti-eIF2α was purchased from Invitrogen; Rabbit anti-actin from Sigma-Aldrich; Mouse anti-Puromycin was obtained from Kerafast, Inc. Mouse anti-G3BP1, mouse anti-GAPDH, rabbit anti-His, and rabbit anti-calnexin were obtained from Abcam, and rabbit anti-p-eIF2α (S52) and Alexa Fluor-conjugated species-specific IgG were purchased from Life Technologies. Rabbit anti-NS7 and rabbit anti-NS5 were manufactured and produced by Invitrogen.

Fritzlar S., Aktepe T.E., Chao Y.W., Kenney N.D., McAllaster M.R., Wilen C.B., White P.A, & Mackenzie J.M. (2019). Mouse Norovirus Infection Arrests Host Cell Translation Uncoupled from the Stress Granule-PKR-eIF2α Axis. mBio, 10(3), e00960-19.

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Immunofluorescence Staining of Polyglutamine Proteins

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Cells were fixed with paraformaldehyde (4% in PBS) for 20 min, followed by permeabilization (0.2% Triton X-100 in PBS, 10 min) and blocking (3% bovine serum albumin in 0.2% Triton X-100 in PBS, 10 min). Cells were incubated in primary antibody for 2 h at room temperature (Mouse anti-polyQ (Millipore, #MAB1574, 1:50), Rabbit anti-HTT (Cell Signaling, #5656, 1:200), Rabbit anti-G3BP1 (MBL, #RN048PW, 1:500), Mouse anti-G3BP1 (Abcam, #56574, 1:500), Rabbit anti-PABP (Abcam, #21060, 1:400)). Then, the cells were washed with 0.2% Triton-X/PBS and incubated with secondary antibody (Alexa Fluor 488 Goat anti-Mouse (ThermoFisher Scientific, #A11029, 1:500), Alexa Fluor 568F(ab′)2 Fragment of Goat Anti-Rabbit IgG (H + L) (ThermoFisher Scientific, #A-21069, 1:500), or Alexa Fluor 594 Goat Anti-Rabbit IgG (H + L) (ThermoFisher Scientific, #A-11037, 1:500)) and Hoechst 33342 (Life Technologies, #H3570, 1:1000) for 1 h at room temperature. PBS and distilled water washes were followed and then the cover slips were mounted on Mowiol (Sigma, #324590).

Gutiérrez-Garcia R., Koyuncu S., Hommen F., Bilican S., Lee H.J., Fatima A, & Vilchez D. (2023). G3BP1-dependent mechanism suppressing protein aggregation in Huntington’s models and its demise upon stress granule assembly. Human Molecular Genetics, 32(10), 1607-1621.

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Immunostaining of G3BP1 and Hedls

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Greenberg E., Hochberg-Laufer H., Blanga S., Kinor N, & Shav-Tal Y. (2019). Cytoplasmic DNA can be detected by RNA fluorescence in situ hybridization. Nucleic Acids Research, 47(18), e109.

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Immunofluorescence Assay of RNA-Binding Proteins

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Cells were grown on coverslips, washed with PBS and fixed for 20 min in 4% PFA. Cells were then permeabilized in 0.5% Triton X-100 for 2.5 min. Cells were washed twice with PBS and blocked with 5% BSA for 20 min cells and immunostained for 1 h with a primary antibody. After three washes with PBS, the cells were incubated for 1 h with secondary fluorescent antibodies. Primary antibodies: mouse anti-G3BP1 (Abcam), rabbit anti-hnRNP A1, rabbit anti-eIF2α, rabbit anti-eIF4B, rabbit anti-CBP80, rabbit anti-U2AF65, rabbit anti-eIF4A3, mouse anti-Y14, mouse anti-Aly, mouse anti-UAP56, rabbit anti-Dbp5, rabbit anti-Nup153, mouse anti-NXF1, rabbit anti-Nup214, rabbit anti-Nup358, mouse anti-Nup62, rat anti-Nup98, rabbit anti-TPR, rabbit anti-THOC5, goat anti-TIA1 (Santa Cruz), mouse anti-CBP20, mouse anti-THOC6, mouse anti-SC35 (Sigma), mouse anti-RACK1 (BD Biosciences). Secondary antibodies: Alexa488-labeled goat anti-mouse/rabbit/rat IgG (Abcam), Alexa488-labeled donkey anti-goat IgG, Alexa594-labeled goat anti-mouse/rabbit. Nuclei were counterstained with Hoechst 33342 (Sigma) and coverslips were mounted in mounting medium. For colocalization analysis, the coloc2 ImageJ plugin was used to calculate Pearson's correlation coefficient (R_r) to measure the colocalization degree. One sample t-test was performed.

Hochberg-Laufer H., Schwed-Gross A., Neugebauer K.M, & Shav-Tal Y. (2019). Uncoupling of nucleo-cytoplasmic RNA export and localization during stress. Nucleic Acids Research, 47(9), 4778-4797.

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