- PA5-35365 - Invitrogen Antibodies GLS antibody

Submitted references Pharmacological Inhibition of Glutaminase 1 Normalized the Metabolic State and CD4+ T Cell Response in Sjogren's Syndrome.

Fu J, Pu Y, Wang B, Li H, Yang X, Xie L, Shi H, Wang Z, Yin J, Zhan T, Shao Y, Chen C, Luo Q, Xu J, Zong Z, Wei X, Xiao W, Yu C, Zheng L
Journal of immunology research 2022;2022:3210200

Quantitative proteomic analysis of enhanced cellular effects of electrochemotherapy with Cisplatin in triple-negative breast cancer cells.

Mittal L, Aryal UK, Camarillo IG, Ferreira RM, Sundararajan R
Scientific reports 2019 Sep 26;9(1):13916

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of GLS in Hela cells using a GLS polyclonal antibody (Product # PA5-35365) followed by detection using a fluorescent conjugated secondary antibody (green). Nuclei were stained with Dapi (blue).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of GLS was performed using 70% confluent log phase Hep G2 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with GLS Polyclonal Antibody (Product # PA5-35365) at 1:50 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Mitochondria (Panel c: red) was stained with Mitotracker Red CMXRos (Product # M7512). Panel d represents the merged image showing mitochondrial localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of GLS was achieved by transfecting Hep G2 cells with GLS specific siRNAs (Silencer® select Product # s5839, s5840). Immunofluorescence analysis was performed using untransfected Hep G2 cells (panels a, d), transfected with non-specific scrambled siRNA (panels b,e) and transfected with GLS specific siRNAs (panel c,f). Cells were fixed, permeabilized, and probed with GLS Polyclonal Antibody (Product # PA5-35365, 1:50 dilution), followed by labelling with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034; 1:2000 dilution) for 45 minutes at room temperature. Nuclei (blue) were stained using ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962) and Rhodamine Phalloidin (Product # R415, 1:300) was used for cytoskeletal F-actin (red) staining. Reduction of mitochondrial GLS expression was observed upon siRNA mediated knockdown (panel c,f), confirming the specificity of the antibody to GLS. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of GLS in HepG2 cells (right) compared to a negative control (left) using a GLS polyclonal antibody (Product # PA5-35365) followed by detection using a FITC-conjugated goat-anti-rabbit secondary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 1 Gls1 was upregulated in SS patients and could be induced upon activation. (a) Expression of Gls1 in circulating CD4+ T cells obtained from healthy individuals and SS patients. n > 25. (b) Immunohistochemical staining of CD4-positive cells and Gls1-positive cells in the labial glands of SS patients. (c) qRT-PCR analysis of Gls1 mRNA expression in murine CD4+ T cells simulated with anti-CD3 and anti-CD28 antibodies in a time-dependent manner. (d) qRT-PCR analysis of Gls1 mRNA expression in human purified circulating CD4+ T cells simulated with anti-CD3 and anti-CD28 antibodies in a time-dependent manner. (e) Representative Western blot pictures of Gls1 protein expression in human purified circulating CD4+ T cells simulated with anti-CD3 and anti-CD28 antibodies. (f) Representative Western blot pictures of Gls1 protein expression in murine splenic CD4+ T cells simulated with anti-CD3 and anti-CD28 antibodies. (g) Representative picture of Gls1 expression and location in murine CD4+ T cells under a confocal microscope. Blue light indicates the position of nuclei; green light indicates the position of the cytoskeleton; red light indicates the position of mitochondria; violet light indicates the position and strength of Gls1. Summary data are presented as ** p < 0.001 with unpaired two-tailed Student's t -tests.

PA5-35365

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