Ab37067

Manufactured by Abcam

Sourced in United States

Ab37067 is a lab equipment product offered by Abcam. It serves a core function, but a detailed description while maintaining an unbiased and factual approach is not available.

Automatically generated - may contain errors

Lab products found in correlation

Ripa assay buffer, by Merck Group (1 mentions) Odyssey clx system, by LI COR (1 mentions) Anti gapdh antibody, by Santa Cruz Biotechnology (1 mentions) Odyssey blocking buffer, by LI COR (1 mentions) Sc 32233, by Santa Cruz Biotechnology (1 mentions) Pvdf membrane, by Bio-Rad (1 mentions) Bradford protein assay, by Bio-Rad (1 mentions) Galectin 3, by Thermo Fisher Scientific (1 mentions) Hif 2α, by R&D Systems (1 mentions) Anti hydroxyproline, by Abcam (1 mentions) β actin, by Cell Signaling Technology (1 mentions) Hif 1α, by BD (1 mentions) Ab30782, by Abcam (1 mentions) Ab50982, by Abcam (1 mentions) Ab6276, by Abcam (1 mentions)

3 protocols using ab37067

Immunoblot Analysis of Collagen Modifiers

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Cells were lysed in Radioimmunoprecipitation (RIPA) assay buffer (Sigma, St. Louis, MO), total protein was extracted, and the protein concentration was measured by Bradford Protein Assay (Bio-Rad). Protein samples were loaded on 10% SDS-PAGE gel and the separated proteins were transferred by electroblotting to polyvinylidene fluoride (PVDF) membranes (Bio-Rad). The membranes were blocked with Odyssey^® Blocking Buffer (LI-COR Biosciences, Lincoln, NE) and incubated with the primary antibody and secondary antibody. Immunolabeling was detected by using Odyssey^® CLx system (LI-COR Biosciences). The following primary antibodies were used: Anti-Hydroxyproline antibody (ab37067, Abcam; 1:1000 dilution); Anti-P4HA1 antibody (ab59497, Abcam; 1:500 dilution); Anti-P4HA2 antibody (A4262, ABclonal Inc.; 1:500 dilution); Anti-GAPDH antibody (sc-32233, Santa Cruz Biotechnology, Inc.; 1:1000 dilution).

Zhu H., Su Y., Wang J, & Wu J.Y. (2023). The role of vesicle trafficking genes in osteoblast differentiation and function. Scientific Reports, 13, 16079.

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Western Blot Analysis of HIF-1α and Related Proteins

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Cells were washed with cold PBS and immediately lysed in 1X SDS-PAGE Loading Buffer (63mM Tris-HCl, pH 6.8, 2% (w/v) SDS, 10% glycerol, 0.01% (w/v) bromophenol blue, and 40mM dithiothreitol). Total cell lysates were separated by SDS-PAGE, transferred to a nitrocellulose membrane and probed with HIF-1α (human: 610958, BD Biosciences, San Jose, CA, USA; mouse: ab2185, Abcam, Cambridge, UK), HIF-2α (AF2997, R&D Systems, Minneapolis, MN, USA), P4HA1 (NBP1-84398, Novus Biologicals, Centennial, CO, USA), P4HA2 (MA5-24599, Thermo Fisher Scientific), anti-hydroxyproline (ab37067, Abcam), Col6A1 (A9236, ABclonal, Woburn, MA, USA), Galectin 3 (PA5-80773, Thermo Fisher Scientific), and β-actin (#4970, Cell Signaling Technology, Danvers, MA, USA) antibodies. Detection was as described previously [23 (link)]. Signal intensity was quantitated by ImageJ (Rasband, W.S., ImageJ, U. S. National Institutes of Health, Bethesda, Maryland, USA, https://imagej.nih.gov/ij/, 1997–2018).

Kaluz S., Zhang Q., Kuranaga Y., Yang H., Osuka S., Bhattacharya D., Devi N.S., Mun J., Wang W., Zhang R., Goodman M.M., Grossniklaus H.E, & Van Meir E.G. (2021). Targeting HIF-activated collagen prolyl 4-hydroxylase expression disrupts collagen deposition and blocks primary and metastatic uveal melanoma growth. Oncogene, 40(33), 5182-5191.

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Comprehensive Immunoblotting Antibody Panel

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The following primary antibodies (Table 1) were used in immunoblotting: HIF-1α (1:1000, 610959, BD), PKM2 (1:1000, 4053 S, CST), PHD3 (1:1000, ab30782, Abcam), hnRNPK (1:1000, sc53620, santa cruz), hnRNPF (1:500, ab50982, abcam), JMJD5 (1:1000, ab10639, abcam), hydroxyproline (1:500, ab37067, abcam), β-actin (1:1000, ab6276,abcam), laminB (1:1000,12255 S,CST), MnSOD (1:1000,611580,BD), HIF-2α(1:1000,59973 S.CST), PKM1(1:1000,15821-1-AP, proteintech).

Zheng F., Chen J., Zhang X., Wang Z., Chen J., Lin X., Huang H., Fu W., Liang J., Wu W., Li B., Yao H., Hu H, & Song E. (2021). The HIF-1α antisense long non-coding RNA drives a positive feedback loop of HIF-1α mediated transactivation and glycolysis. Nature Communications, 12, 1341.

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