Ab93434

Manufactured by Abcam

Sourced in United States, United Kingdom

Ab93434 is a laboratory equipment product. It is a tool designed for research and experimental purposes in a laboratory setting. The core function of this product is to facilitate specific tasks or processes within a laboratory environment.

Automatically generated - may contain errors

Lab products found in correlation

Ab26116, by Abcam (2 mentions) Ab8227, by Abcam (2 mentions) Bca protein assay, by Thermo Fisher Scientific (2 mentions) Ab184714, by Abcam (1 mentions) Ab95950, by Abcam (1 mentions) D2u3t, by Cell Signaling Technology (1 mentions) Image studio lite, by LI COR (1 mentions) Halt protease inhibitor, by Thermo Fisher Scientific (1 mentions) Ab113509, by Abcam (1 mentions) Ab55138, by Abcam (1 mentions) A21285, by Thermo Fisher Scientific (1 mentions) Superfrost slide, by Thermo Fisher Scientific (1 mentions) Axio imager m1 microscope, by Zeiss (1 mentions) Axiocam mrm camera, by Zeiss (1 mentions) Ab21027, by Abcam (1 mentions) Ab32575, by Abcam (1 mentions) Levamisole, by Vector Laboratories (1 mentions) Bx51 microscope, by Zeiss (1 mentions) Vector red alkaline phosphatase substrate solution, by Vector Laboratories (1 mentions) Lsm exciter confocal microscope, by Zeiss (1 mentions)

19 protocols using ab93434

Immunoblotting of Hypoxia Pathway Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

Tissues or cells were lysed on ice in 1× RIPA lysis and extraction buffer (cat. #89900, Thermo Fisher Scientific) with Halt protease inhibitor (cat. #78441, Thermo Fisher Scientific). Protein concentrations were determined using the Pierce BCA Protein Assay Kit (cat. #23225, Thermo Fisher Scientific). Total protein (20 µg) was loaded onto the gels, and blots were developed with primary antibodies against PHD3 (ab184714, Abcam, Cambridge, UK; 1:2000 dilution); HIF1α (D2U3T, Cell Signaling Technology, Danvers, MA, USA; 1:1000 dilution); HIF2α (D9E3, Cell Signaling Technology; 1:1000 dilution); fumarate hydratase (ab95950, Abcam; 1:3000 dilution); EGFR (sc-373746, Santa Cruz Biotechnology, Dallas, TX; 1:1000 dilution). pEGFR (sc-57545, Santa Cruz Biotechnology; 1:200 dilution); glutaminase (GLS1; ab93434, Abcam; 1:1000 dilution), glutaminase 2 (GLS2; NBPI-76544, Novus Biologicals, Littleton, CO, USA; 1:1000 dilution); alanine-serine-cysteine transporter 2 (ASCT2; ABN73, Millipore Sigma, 1:1000 dilution), and β-actin-horseradish peroxidase (5125S or 12262S, Cell Signaling Technology; 1:5000 dilution). Image J (National Institutes of Health, Bethesda, MD, USA) or Image Studio Lite (version 5.2, LI-COR Biosciences, Lincoln, NE, USA) were used to perform densitometry.

Zacharias N.M., Wang L., Maity T., Li L., Millward S.W., Karam J.A., Wood C.G, & Navai N. (2021). Prolyl Hydroxylase 3 Knockdown Accelerates VHL-Mutant Kidney Cancer Growth In Vivo. International Journal of Molecular Sciences, 22(6), 2849.

+ Open protocol

+ Expand

Immunofluorescence Analysis of Corneal Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

Excised mouse eyes or human donor cornea button were fixed in 4% paraformaldehyde in 0.1 mol/L phosphate buffer (pH 7.4) at 4 °C overnight and paraffin embedded. Five-micrometer sections were then mounted on Super Frost slides (Fisher Scientific). The sections were de-paraffinized and hydrated in a graded ethanol series (100%, 95% and 70% and 50% ethanol and ddH₂O for 5 min each) and subject to antigen retrieval in 10 mM Na-citrate, then blocked with 2% BSA in PBS and incubated overnight at 4 °C with 1st antibodies. After washes in PBS, slides were incubated with Fluor-conjugated 2nd antibody for one hour and washed. Sections were mounted with prolong anti-fade mounting reagent with DAPI (Molecular Probes, Life Technologies) and imaged with AxioImager M1 microscope with AxioCam MRm camera (Zeiss).
The following antibodies were used: Rabbit polyclonal anti-Glutaminase (GLS1) antibody 1:200 (ab93434, Abcam); Rabbit anti-GLS2 antibody 1:200 (ab113509, Abcam); Mouse monoclonal anti-GGT1 antibody 1:200 (ab55138, Abcam); Rabbit anti-ZO1 1:200 (402200, Life Technologies); Mouse anti-ZO1 1:200 (339100, Invitrogen); Rabbit anti-Nitrotyrosine 1:200 (A-21285, Thermo Scientific); Secondary Alexa-488 and Alexa-568 antibodies (Molecular Probes) were used at 1:200 concentrations. Mean intensity (MFI) quantification of endothelium was conducted using ImageJ.

Zhang W., Li H., Ogando D.G., Li S., Feng M., Price FW J.r., Tennessen J.M, & Bonanno J.A. (2017). Glutaminolysis is Essential for Energy Production and Ion Transport in Human Corneal Endothelium. EBioMedicine, 16, 292-301.

+ Open protocol

+ Expand

Immunohistochemical Analysis of Mammary Tumors

Check if the same lab product or an alternative is used in the 5 most similar protocols

Mammary tumor sections (5μm) were deparaffinized and high temperature antigen retrieval was performed, followed by blocking in TBS/BSA 5%, 10% donkey serum and exposure to primary antibody and biotinylated secondary antibody (Vectastain ABC kit, Vector Labs) for immunohistochemistry or Alexa 488, 568 and 647-conjugated secondary antibodies (Thermo Fisher Scientific) for immunofluorescence. Primary antibodies against, GLS1 (ab93434; 1/100) and α-SMA (ab32575; 1/1000 or ab21027; 1/300) were purchased from Abcam. A primary antibody against α-SMA (A2547; 1/300) was purchased from Sigma. A primary antibody against SLC1A3 (sc-7757; 1/100), was purchased from Santa Cruz Biotechnology. A primary antibody against PCNA (13–3900, 1/100) was purchased from Thermo Fisher Scientific. In most cases, color development was achieved by adding streptavidin biotinylated alkaline phosphatase complex (Vector Labs) followed by Vector Red alkaline phosphatase substrate solution (Vector Labs). Levamisole was added to block endogenous alkaline phosphatase activity (Vector Labs). Pictures were obtained using an Olympus Bx51 microscope or ZEISS LSM Exciter confocal microscope. Intensity of staining was quantified using ImageJ software (NIH). All measurements were performed blinded to condition

Bertero T., Oldham W.M., Grasset E.M., Bourget I., Boulter E., Pisano S., Hofman P., Bellvert F., Meneguzzi G., Bulavin D.V., Estrach S., Feral C.C., Chan S.Y., Bozec A, & Gaggioli C. (2018). Tumor-stroma mechanics coordinate amino acid availability to sustain tumor growth and malignancy. Cell metabolism, 29(1), 124-140.e10.

+ Open protocol

+ Expand

Protein Expression Analysis in ESCs

Check if the same lab product or an alternative is used in the 5 most similar protocols

Protein extraction was prepared from ESCs using RIPA buffer (Beyotime Biotechnology) with proteinase and phosphatase inhibitors. After being quantified by the BCA assay kit (Thermo Scientific), equal amounts of proteins were separated by SDS-PAGE gel electrophoresis and transferred to polyvinylidene difluoride membranes (Merck Millipore). Membranes were blocked in 5% BSA (Sigma) at room temperature for 1 h, followed by incubation with primary antibodies against GAPDH (1:10000; Proteintech, 60004-1-Ig), GLS1 (1:1000; Abcam, ab93434), Collagen I (1:1000; Abcam, ab34710), α-SMA (1:1000; Abcam, ab7817), MFN1(1:1000; Abcam, ab57602), MFN2(1:1000; Abcam, ab56889), DRP1(1:1000; Abcam, ab56788), PINK1(1:1000; Cell Signaling Technology, 6946), p-p70-S6K (1:1000; Cell Signaling Technology, 9234), p-4E-BP1 (1:1000; Cell Signaling Technology, 2855) and p-S6 (1:1000; Cell Signaling Technology, 4858) overnight at 4 °C. The membrane was then incubated with HRP-conjugated goat anti-rabbit (1:10000; Proteintech, SA00001-2) or goat anti-mouse (1:10000, Proteintech, SA00001-1) secondary antibodies at room temperature for 1 h. Pageruler (ThermoFisher, 26,616) was used as the molecular marker. Protein expression was visualized with an ECL detection kit (Sigma-Aldrich). GAPDH was used as the loading control.

Chen P., Ye C., Huang Y., Xu B., Wu T., Dong Y., Jin Y., Zhao L., Hu C., Mao J, & Wu R. (2024). Glutaminolysis regulates endometrial fibrosis in intrauterine adhesion via modulating mitochondrial function. Biological Research, 57, 13.

+ Open protocol

+ Expand

Co-localization of GLS1 and α-SMA in TGF-β1-treated ESCs

Check if the same lab product or an alternative is used in the 5 most similar protocols

Immunofluorescence were performed in TGF-β1-treated ESCs to detect the co-localization of GLS1 and α-SMA. ESCs were seeded on 35 mm confocal dishes (Cellvis) at a density of 1 × 10⁴ cells/mL. After treatments, cells were fixed with 4% PFA for 15 min, permeabilized with 0.2% Triton X for 10 min and then blocked with 5% BSA for 45 min at room temperature. Primary antibodies α-SMA (1:200; Abcam, ab7818), GLS1 (1:200; Abcam, ab93434) were used for overnight incubation at 4 °C. DyLight488-conjugated goat anti-rabbit antibody (1:200; Multisciences, GAR4882) and DyLight594-conjugated goat anti-mouse antibody (1:200; Multisciences, GAM5942) were used for detection. DAPI was used to stain cell nucleus, and the stained ESCs were observed by an Olympus FV1200 confocal microscope (Olympus).

+ Open protocol

+ Expand

Glutaminase Isoforms in AML3 Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

The OCI-AML3 cells stably transfected with pHUSH luciferase control, pHUSH puro-KGA, pHUSH puro-GAC, or pHUSH puro-D3 shRNA were grown, collected, washed in PBS, and pelleted before lysing with RIPA buffer (PIERCE) containing protease and phosphatase inhibitors (Roche). After 30 min incubation on ice, cell lysates were subject to sonication (~2 min) and centrifugation (12,000 rpm x 10 min, 4° C). The supernatant was collected and the protein concentration quantified using the BCA Protein Assay (Thermo Scientific). Western blot analysis performed according to standard procedures. Antibodies used in western analyses: GLS (ab93434, Abcam), GAPDH (MAB374, MilliporeSigma), LDH (2012S, Cell Signal Technology), α-tubulin (ab7291, Abcam); and donkey anti-mouse antibody IRDye 800CW (C61116–02, LI-COR), donkey anti-rabbit antibody (C70601–05, LI-COR). GLS (ab93434) recognizes both the KGA and GAC isotypes but not GLS2 [28 (link),29 (link)].

Zacharias N.M., Baran N., Shanmugavelandy S.S., Lee J., Lujan J.V., Dutta P., Millward S.W., Cai T., Wood C.G., Piwnica-Worms D., Konopleva M, & Bhattacharya P.K. (2019). Assessing Metabolic Intervention with a Glutaminase Inhibitor in Real-time by Hyperpolarized Magnetic Resonance in Acute Myeloid Leukemia. Molecular cancer therapeutics, 18(11), 1937-1946.

+ Open protocol

+ Expand

Profiling Hepatocellular Carcinoma Biomarkers

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Tissue microarrays containing 75 paired hepatocellular carcinoma tissues and para-tumor tissues were purchased from Shanghai Outdo Biotech Company (HLiv-HCC150PG-01). The detailed information of the tissue microarrays could be downloaded from website (Error! Hyperlink reference not valid.). IHC staining was performed according to standard protocol using anti-SREBF2 (ab30682, 1:200), anti-SLC1A5 (ab187692, 1:100) and anti-GLS (ab93434, 1:200) from Abcam. The expression of each protein in the tissue was evaluated independently by three pathologists and scored according to staining intensity and percentage as previously described [46 (link)].

Kong Y., Wu M., Wan X., Sun M., Zhang Y., Wu Z., Li C., Liang X., Gao L., Ma C, & Yue X. (2023). Lipophagy-mediated cholesterol synthesis inhibition is required for the survival of hepatocellular carcinoma under glutamine deprivation. Redox Biology, 63, 102732.

+ Open protocol

+ Expand

Neurotransmitter Receptor Immunostaining

Check if the same lab product or an alternative is used in the 5 most similar protocols

Antibodies included those against GLS (1:25 for immunohistochemistry, 1:100 for immunocytochemistry, ab93434, Abcam), GRINA (1:50 for immunohistochemistry, 1:100 for immunocytochemistry, ab216953, Abcam), GLUR3 (1:50 for immunohistochemistry, 1:100 for immunocytochemistry, ab232887, Abcam), mGluR1 (1:50 for immunohistochemistry, 1:100 for immunocytochemistry, NB110-39033SS, Novus), mGluR5 (1:50 for immunohistochemistry, 1:100 for immunocytochemistry, ab76316, Abcam), VGLUT1 (1:50 for immunohistochemistry, 48-2400, Invitrogen; 1:100 for immunocytochemistry, 135 303, Synaptic Systems), UNC13B (1:50 for immunohistochemistry, 1:100 for immunocytochemistry, TA308990, Origene), SNAP25 (1:50 for immunohistochemistry, GTX113839, Gene Tex; 1:100 for immunocytochemistry, ab31281, Abcam), GAD1 (1:50 for immunohistochemistry, 1:100 for immunocytochemistry, ab26116, Abcam), SYT11 (1:50 for immunohistochemistry, 1:100 for immunocytochemistry, PA5-96970, Invitrogen), CAST (1:50 for immunohistochemistry, 1:100 for immunocytochemistry, PA5-87352, Invitrogen), and HCN4 (1:50 for immunohistochemistry, 1:100 for immunocytochemistry, SAB5200035, Sigma; SMC-320, StressMarq Biosciences). Alexa Fluor conjugated secondary antibodies were purchased from Abcam.

Liang D., Xue Z., Xue J., Xie D., Xiong K., Zhou H., Zhang F., Su X., Wang G., Zou Q., Liu Y., Yang J., Ma H., Peng L., Zeng C., Li G., Wang L, & Chen Y.H. (2021). Sinoatrial node pacemaker cells share dominant biological properties with glutamatergic neurons. Protein & Cell, 12(7), 545-556.

+ Open protocol

+ Expand

Western Blot Protein Detection

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Western blots were carried out as previously described [25 (link)]. Primary antibodies for protein detection included: GLS1 (ab93434, Abcam), PARP1 (F-2, Santa Cruz), PAR (Trevigen, Gaithersburg, MD), Actin (C-2, Santa Cruz), small subunit calpain (EPR3324, Abcam), and 53BP1 (Bethyl Laboratories).

Chakrabarti G., Moore Z.R., Luo X., Ilcheva M., Ali A., Padanad M., Zhou Y., Xie Y., Burma S., Scaglioni P.P., Cantley L.C., DeBerardinis R.J., Kimmelman A.C., Lyssiotis C.A, & Boothman D.A. (2015). Targeting glutamine metabolism sensitizes pancreatic cancer to PARP-driven metabolic catastrophe induced by ß-lapachone. Cancer & Metabolism, 3, 12.

+ Open protocol

+ Expand

Phosphorylation of Glutaminase GAC

Check if the same lab product or an alternative is used in the 5 most similar protocols

Protein extracts were prepared using NP-40 lysis buffer containing phosphatase and protease inhibitors and the cell lysates were then subjected to Laemmli SDS-PAGE followed by immunoblot using indicated antibodies. The anti-GAC antibody was purchased from Abcam (ab93434). The anti-phospho-serine 314 specific polyclonal antibody against GAC (GAC-pS314) was made by Shanghai Genomics Inc (antigen sequence: YVGKEPS(p)GLRFNK-C; immunogen: Peptide-KLH conjugated). Isoelectric focusing was conducted as previously described^{44 (link)} followed by immunoblot with the anti-V5 antibody.

Han T., Zhan W., Gan M., Liu F., Yu B., Chin Y.E, & Wang J.B. (2018). Phosphorylation of glutaminase by PKCε is essential for its enzymatic activity and critically contributes to tumorigenesis. Cell Research, 28(6), 655-669.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!