Tissue lysates were analyzed using automated Simple Wes system (Protein Simple) with 12–230 kDa Wes Separation Module capillary cartridges (SM-W004, Protein Simple). Anti- mouse (DM-002, Protein Simple), anti-rabbit (DM-001, Protein Simple), anti-goat (DM-006, Protein Simple) detection modules or F(ab’)2 anti-Rat IgG (H + L)-HRPO (1:20, 112-036-062, Jackson Immuno Research) were used, depending on host species of the primary antibodies. The following primary antibodies were used: LDLR (1:50, PAB8804, Abnova), SERPINA1A (1:20, MAB7690, R&D Systems), PCSK9 (1:10, AF3985, R&D Systems) and β-actin (1:20, NB600-501, Novus Biologicals). Specificity of PCSK9 detection was confirmed using recombinant mouse PCSK9 protein (9258-SE-022, R&D Systems). Compass software version 6.0.0 (Protein Simple) was used to analyze the data. Area under the peak of the protein of interest was measured and normalized with respect to the β-actin area under the peak.
Pab8804
Manufactured by Abnova
PAB8804 is a laboratory equipment product manufactured by Abnova. It is designed to perform a specific function within a laboratory setting, but a detailed and unbiased description of its core function cannot be provided while maintaining the requested level of conciseness and objectivity.
Automatically generated - may contain errors
Lab products found in correlation
Complete mini protease inhibitor cocktail, by Merck Group (2 mentions)
Af2474 primary antibody, by R&D Systems (2 mentions)
Af3985, by R&D Systems (1 mentions)
Nb600 501, by Novus Biologicals (1 mentions)
Cell lysis buffer 9803, by Cell Signaling Technology (1 mentions)
Goat anti mouse igg h l hrp conjugate, by Bio-Rad (1 mentions)
Image lab 3, by Bio-Rad (1 mentions)
A5441, by Merck Group (1 mentions)
Ab4047, by Abcam (1 mentions)
Ab2900, by Abcam (1 mentions)
Sc 459, by Santa Cruz Biotechnology (1 mentions)
Goat anti rabbit igg h l horseradish peroxidase hrp conjugate, by Bio-Rad (1 mentions)
4 protocols using pab8804
1
Quantification of LDLR, SERPINA1A, and PCSK9
2
LDLR Immunoblot Analysis in Hepatocytes
3
LDLR Immunoblot Protocol with Simple Western
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Hepatocytes were washed twice with phosphate-buffered saline (PBS), and cellular proteins were extracted in 100-μl cell lysis buffer 9803 (Cell Signaling Technology, Saint Quentin en Yvelines, France) supplemented with complete mini-protease inhibitor cocktail (Sigma-Aldrich, St. Louis, Missouri). LDLR immunoblot analyses were performed with a rabbit antihuman-LDLR polyclonal antibody PAB8804 (Abnova, Walnut, California) on a Sally Sue Simple Western device, using the 66-440 kDa separation master kit PS-MK12 (Protein Simple, San Jose, California), and normalized to transferrin receptor expression assessed using the AF2474 primary antibody (R&D Systems, Lille, France).
4
COMMD1 and LDLR Protein Detection Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
In the experimental procedures described, the following antibodies were used: rabbit polyclonal antibody against COMMD1 (11938-1-AP, Proteintech Group, 1:1,000); rabbit polyclonal antibody against LDLR (PAB8804, Abnova, 1:1,000); rabbit polyclonal antibody against GST (Z-5) (sc-459, Santa Cruz Biotechnology, 1:5,000); goat anti-rabbit IgG (H+L)-horseradish peroxidase (HRP) conjugate (170-6515, Bio-Rad Laboratories, 1:10,000); goat anti-mouse IgG (H+L)-HRP conjugate (170-6516, Bio-Rad Laboratories, 1:10,000); mouse anti-β-actin (A5441, Sigma-Aldrich, 1:5,000); rabbit anti-tubulin (AB4047, Abcam, 1:2,000); rabbit anti-Rab11 (700184, Invitrogen, 1:1,000); rabbit anti-EEA-1 (AB2900, Abcam, 1:1,000); rabbit anti-CCDC22 (16636-1-AP, Proteintech, 1:2,000); and rabbit anti-CCDC93 (20861-1-AP, Proteintech, 1:5,000). Rabbit polyclonal antibodies against WASH1, FAM21 and VPS35 were previously described1 (link)5 (link). Anti-apoB100 (1:1,000) and rabbit anti-apoA1 (1:1,000) antibodies were a gift from A.K. Groen. Rabbit polyclonal antibody against ARH (1:1,000) was a gift from H.H. Hobbs. Densitometry analysis of western blot bands was performed using Image Lab 3.0.1 software (Bio-Rad Laboratories). Images have been cropped for presentation. Full-size images are presented in Supplementary Fig. 12 .
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!