Ab94571

Manufactured by Abcam

Sourced in United States

Ab94571 is a lab equipment product manufactured by Abcam. It is a device used for performing laboratory experiments and analyses. The core function of this product is to facilitate the completion of various tasks within a controlled laboratory environment.

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Lab products found in correlation

Bca protein assay, by Thermo Fisher Scientific (4 mentions) Ab55271, by Abcam (3 mentions) Ab58641, by Abcam (2 mentions) Pdhα pser293, by Merck Group (2 mentions) Aif sc 13116, by Santa Cruz Biotechnology (2 mentions) Ctp sc 86392, by Santa Cruz Biotechnology (2 mentions) Gapdh ab2302, by Merck Group (2 mentions) Hif 1α, by Cell Signaling Technology (2 mentions) Actin, by Merck Group (2 mentions) Ab2430, by Abcam (1 mentions) Ab868, by Abcam (1 mentions) Deoxycholate, by Merck Group (1 mentions) Ab108630, by Abcam (1 mentions) Ab131442, by Abcam (1 mentions) Hybond ecl membrane, by GE Healthcare (1 mentions) H 300, by Santa Cruz Biotechnology (1 mentions) Igepal, by MP Biomedicals (1 mentions) Complete mini protease inhibitor cocktail, by Roche (1 mentions) Phosphatase inhibitor cocktail, by Roche (1 mentions) Dia h09, by Dianova (1 mentions)

4 protocols using ab94571

Comprehensive Protein Expression Analysis

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Confluent cells were lysed in 50 mM Tris-HCl (pH 8.0), 150 mM NaCl, 1 mM EDTA, 1% Igepal (MP Biomedicals), 0.5% deoxycholate (Sigma Aldrich), and 0.1% SDS with Complete Mini protease inhibitor cocktail and phosphatase inhibitor cocktail (Roche) and stored at −80°C until analyzed. A431 control cell lysate (ab7909) was purchased from Abcam. Aliquots were run on 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis gels and blotted to Hybond-ECL membranes (GE Healthcare). Proteins were detected with primary antibodies against CAR (1:500, H-300, Santa Cruz Biotechnology Inc.), CD46 (1:500), DSG-2 (1:500), S100 (1:500, ab868, Abcam), p53 (1:500, ab131442, Abcam), EGFR (1:200, ab2430, Abcam), isocitrate dehydrogenase (wild-type IDH1, 1:600, ab94571, Abcam), IDH1 R132H (the most common IDH1 mutant, 1:500, DIA-H09, Dianova, USA), MGMT (1:1000, ab108630, Abcam), and β-actin (1:2500, C4, Santa Cruz Biotechnology). Blots were developed with anti-rabbit-Cy5 and anti-mouse-Cy3 secondary antibodies of Amersham ECL Plex Western blotting system (GE Healthcare).

Niittykoski M., von und zu Fraunberg M., Martikainen M., Rauramaa T., Immonen A., Koponen S., Leinonen V., Vähä-Koskela M., Zhang Q., Kühnel F., Mei Y.F., Ylä-Herttuala S., Jääskeläinen J.E, & Hinkkanen A. (2017). Immunohistochemical Characterization and Sensitivity to Human Adenovirus Serotypes 3, 5, and 11p of New Cell Lines Derived from Human Diffuse Grade II to IV Gliomas. Translational Oncology, 10(5), 772-779.

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Quantifying Metabolic Enzyme Profiles

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Whole cells/spheroids or mitochondrial lysates were prepared in RIPA buffer and quantified using the BCA Protein Assay (Thermo Scientific). Proteins were separated on 4–20% SDS-PAGE gels, transferred to PVDF membranes, and probed with antibodies against IDH1 (ab94571), IDH2 (ab55271), IDH3 (ab58641) from Abcam, PDHα (#459400, Theromo), PDHα-pSer293 (AP1062), GAPDH (AB2302) from Millipore, PDK1 (#3820), Hif1α(#3716) from Cell Signaling, CTP (sc-86392), AIF (sc-13116) from Santa Cruz Biotechnology and Actin (A3853, Sigma).

Jiang L., Shestov A.A., Swain P., Yang C., Parker S.J., Wang Q.A., Terada L.S., Adams N.D., McCabe M.T., Pietrak B., Schmidt S., Metallo C.M., Dranka B.P., Schwartz B, & DeBerardinis R.J. (2016). Reductive carboxylation supports redox homeostasis during anchorage-independent growth. Nature, 532(7598), 255-258.

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Metabolic Enzyme Profiling in Cellular Systems

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Antibodies to citrate synthetase (ab96600), GAPDH (ab8245), IDH1 (ab94571), and IDH2 (ab55271) were from Abcam. Anti-Tubulin Antibody (MAB1637) was from EMD Millipore. IRDye 680RD goat anti-rabbit (926–68071) and IRDye 800CW goat anti-mouse (926–32210) secondary antibodies were from LI-COR. Primary and secondary antibodies were used at 1:1000 and 1:15,000 dilutions, respectively. Digitonin (D141), α-ketoglutaric acid (K3752), citric acid (C7129), malic acid (M1000), and sodium dichloroacetate (347795) were purchased from Sigma; glutamic acid (41–217–25)—from Biological Industries. [U-¹³C]-glutamine (CLM-1822-H-0.1), [1-¹³C]-glutamine (CLM-3612), and [U-¹³C]-glucose (CLM-1396-1) were purchased from Cambridge Isotope Laboratories.

Lee W.D., Mukha D., Aizenshtein E, & Shlomi T. (2019). Spatial-fluxomics provides a subcellular-compartmentalized view of reductive glutamine metabolism in cancer cells. Nature Communications, 10, 1351.

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Quantifying Metabolic Enzyme Profiles

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