Impdh2

Manufactured by Abcam

IMPDH2 is an enzyme that catalyzes the conversion of inosine monophosphate (IMP) to xanthosine monophosphate (XMP) in the de novo purine biosynthesis pathway. It plays a crucial role in the regulation of cellular guanine nucleotide levels.

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Lab products found in correlation

Tubulin, by Proteintech (1 mentions) Impdh2, by Proteintech (1 mentions) Cdc42, by Cell Signaling Technology (1 mentions) Pierce anti c myc magnetic beads, by Thermo Fisher Scientific (1 mentions) Itgb1, by Proteintech (1 mentions) Impdh1, by Proteintech (1 mentions) Gk10008, by GLPBIO (1 mentions) β actin, by Proteintech (1 mentions) Phenyl methyl sulfonyl fluoride (pmsf), by Beyotime (1 mentions) Gapdh, by Proteintech (1 mentions) Radioimmunoprecipitation assay (ripa), by Beyotime (1 mentions) Rpa32 rpa2, by Abcam (1 mentions) Protease inhibitor cocktail, by Merck Group (1 mentions) Phosphatase inhibitor cocktail 2, by Merck Group (1 mentions)

3 protocols using impdh2

Immunoprecipitation and Immunoblotting Assays

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Immunoprecipitation was performed using the following antibodies: IMPDH2 (Abcam, ab129165), RAC1 (Santa Cruz Biotechnology, sc-514583), Pierce™ Anti-c-Myc Magnetic Beads (Thermo Fisher Scientific, 88842). Immunoblotting was performed with the following antibodies: RAC1 (Proteintech, 24072-1-AP 1:500), GMPS (Abcam, ab228716 1:1000), IMPDH2 (Proteintech, 67663-1 1:500), Tubulin (Proteintech, HRP-66031 1:1000), GMPR (Proteintech, 15683-1-AP 1:1000), NME1 (Proteintech, 11086-2-AP 1:1000), ITGB1 (Proteintech, 26918-1-AP 1:1000), KLF9 ((Santa Cruz Biotechnology, sc-376422 1:250), RHOA (Cell Signaling, #2117, 1:500), CDC42 (Cell Signaling, #8747, 1:200), and RAS (Cell Signaling, #8832, 1:500).

Bianchi-Smiraglia A., Wolff D.W., Marston D.J., Deng Z., Han Z., Moparthy S., Wombacher R.M., Mussell A.L., Shen S., Chen J., Yun D.H., O’Brien Cox A., Furdui C.M., Hurley E., Feltri M.L., Qu J., Hollis T., Kengne J.B., Fongang B., Sousa R.J., Kandel M.E., Kandel E.S., Hahn K.M, & Nikiforov M.A. (2021). Regulation of local GTP availability controls RAC1 activity and cell invasion. Nature Communications, 12, 6091.

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Bladder Cancer Protein Expression Analysis

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RIPA (Beyotime#P0013B) supplemented with phosphatase inhibitor II (ZHHC#PL026) and PMSF (ZHHC#PL012) were used to lyse bladder cancer cells. Protein concentration was determined by BCA (Yamei#ZJ101L) and then separated into 10% SDS-PAGE gels with antibodies for bound protein and ECL blotting system (GLPbio#GK10008) to detect the bands. Primary antibodies were used in the experiments included: β-actin (1:2000; Proteintech#81115-1-RR); GAPDH (1:2000; Proteintech#10494-1-AP); IMPDH1 (1:3000; Proteintech#22092-1-AP); IMPDH2 (1:4000; Abcam#ab131158); TWIST1 (1:70; Santa cruz#sc-81417/ 1:1000; Proteintech#25465-1-AP); NFIL3 (1:150; Santa cruz#sc-74415); IKZF1 (1:150; Santa cruz#sc-398265); TEAD2 (1:150; Santa cruz#sc-81397); GLI2 (1:150; Santa cruz#sc-271786); PRRX1 (1:150; Santa cruz#sc-293386); FLI1 (1:150; Santa cruz#sc-365294); MMP9 (1:150; Santa cruz#sc-13520); PCNA (1:200; Santa cruz#sc-56); MMP2 (1:150; Santa cruz#sc-13595); EMMPRIN (1:150; Santa cruz#sc-21746).

Liu S.S., Li J.S., Xue M., Wu W.J., Li X, & Chen W. (2023). LncRNA UCA1 Participates in De Novo Synthesis of Guanine Nucleotides in Bladder Cancer by Recruiting TWIST1 to Increase IMPDH1/2. International Journal of Biological Sciences, 19(8), 2599-2612.

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Comprehensive Protein Analysis Protocol

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Cells were lysed in buffer containing 20 mM Tris pH 7.5, 140 mM NaCl, 1 mM EDTA, 10% glycerol, 1% Triton X-100, 1 mM DTT, 50 mM NaF, protease inhibitor cocktail (Sigma #P8340) and phosphatase inhibitor cocktail #2 (Sigma #P5726) and #3 (Sigma #P0044) used at 1:100. Western blots were performed using the following antibodies at 1:1000 dilution unless otherwise indicated: phospho-p70 S6 Kinase T389 (CST #9234), p70 S6 Kinase (CST #2708), 4E-BP1 (CST #9644), TSC2 (CST #4308 and #3612), GAPDH (CST #5174), IMPDH1 (Abcam #ab137120), IMPDH2 (Abcam #ab131158), Cleaved Caspase-3 D175 (CST #9664), PARP (CST #9542), phospho-AMPKα T172 (CST #2535), AMPKα (CST #2532), phospho-Raptor S792 (CST #2083), Raptor (CST #2280), ADK (Santa Cruz #sc-514588, 1:500), phospho-Chk1 S345 (CST #2348), Chk1 (CST #2360), RPA32/RPA2 (Abcam #ab76420, 1:10000), phospho-Histone H2A.X S139 (CST #9718), Histone H2A.X (CST #7631), and UBF (Santa Cruz #sc9131).

Valvezan A.J., Turner M., Belaid A., Lam H.C., Miller S.K., McNamara M.C., Baglini C., Housden B.E., Perrimon N., Kwiatkowski D.J., Asara J.M., Henske E.P, & Manning B.D. (2017). mTORC1 couples nucleotide synthesis to nucleotide demand resulting in a targetable metabolic vulnerability. Cancer cell, 32(5), 624-638.e5.

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