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).
Anti hydroxyproline
Manufactured by Abcam
Sourced in United States
Anti-hydroxyproline is an antibody that binds to the amino acid hydroxyproline. Hydroxyproline is a non-standard amino acid found in collagen and other proteins. This antibody can be used to detect and quantify the presence of hydroxyproline in various biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Ab37067, by Abcam (1 mentions)
Galectin 3, by Thermo Fisher Scientific (1 mentions)
Hif 2α, by R&D Systems (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
Hif 1α, by BD (1 mentions)
β actin, by Novus Biologicals (1 mentions)
Protein a agarose beads, by Novus Biologicals (1 mentions)
Anti phd2, by Novus Biologicals (1 mentions)
Anti phd3, by Novus Biologicals (1 mentions)
Anti flag, by Merck Group (1 mentions)
Flag tag (flag), by Merck Group (1 mentions)
2 protocols using anti hydroxyproline
1
Western Blot Analysis of HIF-1α and Related Proteins
2
Immunoprecipitation and Immunoblotting
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were lysed in modified radioimmunoprecipitation assay buffer, and WCLs were incubated overnight with anti-FLAG (Sigma-Aldrich, St. Louis, MO), anti-PHD2 (Novus Biologicals, Littleton, CO), anti-PHD3 (Novus Biologicals), or anti-hydroxyproline (Abcam) antibody in the presence of protein A–agarose beads (Novus Biologicals). After three washes, the bound proteins were fractionated by SDS–PAGE and analyzed by immunoblot assays using antibodies against the following proteins or epitope tag: PHD2, PHD3, β-actin, PKM2 (Novus Biologicals), V5 (Invitrogen, Carlsbad, CA), or FLAG (Sigma-Aldrich).
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?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!