Preparation of heparin media and analyses of EL overexpression were performed as described [22 (link)]. HDL-associated proteins (10 μg) and cell lysates (30 μg) were separated by 12% SDS-PAGE at 175 V for 90 min. Separated proteins were transferred to polyvinylidene difluoride (PVDF) membranes (Carl Roth, Karlsruhe, Germany) with blotting buffer (Tris, glycine, EDTA, sodium azide) at 150 mA for 90 min. Membranes were blocked at RT in 10% skim milk for 2 h followed by overnight incubation at 4 °C with antibodies specific for: human PON1 (Abcam, ab24261, Cambridge, UK), human apoA-I (Novus biological, NB100–65491, Littleton, CO, USA, or Academy biomedical, 11A-G2b, Houston, TX, USA), human LCAT (Novus biological, Littleton, CO, USA), human PAF-AH (Cayman chemical, 160603, Ann Arbor, MI, USA), α-tubulin (Cell signaling technology, 11H10, Leiden, Netherlands) and albumin (Abcam, ab 83465, Cambridge, UK). After washing and incubation with the appropriate secondary antibody (Dako, Vienna, Austria), protein signals were visualized by incubation with Pierce ECL Western Blotting Substrate (Thermo Fisher Scientific, Schwerte, Germany) using the ChemiDoc system (Bio-Rad Laboratories, Vienna, Austria).
Human paf ah
Manufactured by Cayman Chemical
Sourced in United States
Human PAF-AH is a recombinant enzyme that catalyzes the hydrolysis of platelet-activating factor (PAF). It is a key regulator of PAF levels in the body.
Automatically generated - may contain errors
2 protocols using human paf ah
1
Western Blot Analysis of HDL-Associated Proteins
2
Western Blot Analysis of HDL-Associated Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Preparation of heparin media and analyses of EL overexpression were performed as described [22 (link)]. HDL-associated proteins (10 μg) and cell lysates (30 μg) were separated by 12% SDS-PAGE at 175 V for 90 min. Separated proteins were transferred to polyvinylidene difluoride (PVDF) membranes (Carl Roth, Karlsruhe, Germany) with blotting buffer (Tris, glycine, EDTA, sodium azide) at 150 mA for 90 min. Membranes were blocked at RT in 10% skim milk for 2 h followed by overnight incubation at 4 °C with antibodies specific for: human PON1 (Abcam, ab24261, Cambridge, UK), human apoA-I (Novus biological, NB100–65491, Littleton, CO, USA, or Academy biomedical, 11A-G2b, Houston, TX, USA), human LCAT (Novus biological, Littleton, CO, USA), human PAF-AH (Cayman chemical, 160603, Ann Arbor, MI, USA), α-tubulin (Cell signaling technology, 11H10, Leiden, Netherlands) and albumin (Abcam, ab 83465, Cambridge, UK). After washing and incubation with the appropriate secondary antibody (Dako, Vienna, Austria), protein signals were visualized by incubation with Pierce ECL Western Blotting Substrate (Thermo Fisher Scientific, Schwerte, Germany) using the ChemiDoc system (Bio-Rad Laboratories, Vienna, Austria).
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!