Ab110171

Manufactured by Abcam

Ab110171 is an antibody that recognizes the Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) protein. GAPDH is a key enzyme involved in glycolysis, the metabolic pathway that converts glucose into energy. This antibody can be used for the detection and quantification of GAPDH in various applications such as Western blotting, immunohistochemistry, and ELISA.

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Lab products found in correlation

Ab83431, by Abcam (1 mentions) Mak183, by Merck Group (1 mentions) Mak189, by Merck Group (1 mentions) Ab26300, by Abcam (1 mentions) Anti dna2, by Thermo Fisher Scientific (1 mentions) Anti cox 4, by Cell Signaling Technology (1 mentions) Anti vinculin, by Merck Group (1 mentions) Nuclear cytosol fractionation kit, by Abcam (1 mentions)

2 protocols using ab110171

Mitochondrial Enzyme Activity Assays

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Intracellular αKGDH concentration was measured using a commercial kit (ab83431; Abcam). PDH and α-KGDH activities were determined in isolated mitochondria (ab110171; Abcam) using colorimetric enzymatic activity assays (MAK183 and MAK189, respectively; Sigma).

Paredes F., Sheldon K., Lassègue B., Williams H.C., Faidley E.A., Benavides G.A., Torres G., Sanhueza-Olivares F., Yeligar S.M., Griendling K.K., Darley-Usmar V, & San Martin A. (2018). Poldip2 is an oxygen-sensitive protein that controls PDH and αKGDH lipoylation and activation to support metabolic adaptation in hypoxia and cancer. Proceedings of the National Academy of Sciences of the United States of America, 115(8), 1789-1794.

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Isolation and Fractionation of Mitochondrial and Nuclear Proteins

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A mitochondria isolation kit (Abcam, ab110171) was used to prepare the large organelles/debris and intact mitochondria fractions from JJN3 cells. Briefly, cell pellets were frozen and thawed to weaken cell membranes. Cell pellets were resuspended in the extraction buffer and homogenized following the manufacturer’s procedures. After the last centrifugation step of mitochondrial isolation, the supernatants were collected for further nuclear isolation using the nuclear extraction buffer from a nuclear/cytosol fractionation kit (Biovision; K269) following the manufacturer’s procedures. Mitochondrial and nuclear proteins were quantified using the Qubit Protein Assay kit. WB analysis was performed using the following primary antibodies: anti-DNA2 (Invitrogen, PA5-8167, dilution 1:1000), anti-vinculin (Sigma, V9131, dilution 1:2000), anti-COX IV (Cell Signaling, 4850 S, dilution 1:1000), and anti-Lamin A (Abcam, ab26300, dilution 1:1000).

Thongon N., Ma F., Baran N., Lockyer P., Liu J., Jackson C., Rose A., Furudate K., Wildeman B., Marchesini M., Marchica V., Storti P., Todaro G., Ganan-Gomez I., Adema V., Rodriguez-Sevilla J.J., Qing Y., Ha M.J., Fonseca R., Stein C., Class C., Tan L., Attanasio S., Garcia-Manero G., Giuliani N., Berrios Nolasco D., Santoni A., Cerchione C., Bueso-Ramos C., Konopleva M., Lorenzi P., Takahashi K., Manasanch E., Sammarelli G., Kanagal-Shamanna R., Viale A., Chesi M, & Colla S. (2024). Targeting DNA2 overcomes metabolic reprogramming in multiple myeloma. Nature Communications, 15, 1203.

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