Murine tissues were homogenized using metal beads in a Tissue Lyser (Qiagen) as previously described (22 (link)) and cells were lysed in RIPA lysis buffer. 30–50 μg of total protein was resolved per lane by SDS-PAGE and transferred to a 0.45 mm nitrocellulose membrane. Blots were probed with antibodies against β-actin or anti-Ubiquitin (Sigma-Aldrich), p62 (Progen), phosphorylated (Ser 240/244) and total S6 (Cell Signaling), LC3 I/II (Novus Biologicals), GRK2, GAPDH and α-tubulin (Santa Cruz Biotechnology). Immunoreactive bands were visualized using enhanced chemiluminescence (ECL; Amersham Biosciences, Buckinghamshire, UK) or the Odyssey Infrared Imaging System (Li-Cor Biosciences). Films were scanned with a GS-700 Imaging Densitometer and analyzed with Quantity One Software (Bio-Rad), or using an Odyssey Classic reader and the Odyssey software package 3.0 (Li-Cor Biosciences).
Gapdh and α tubulin
Manufactured by Santa Cruz Biotechnology
GAPDH (Glyceraldehyde 3-phosphate dehydrogenase) and α-tubulin are proteins commonly used as loading controls in various biological applications. GAPDH is an enzyme involved in the glycolytic pathway, while α-tubulin is a structural component of microtubules. These proteins are often used as internal standards to normalize the expression levels of target proteins in Western blotting, immunohistochemistry, and other techniques.
Automatically generated - may contain errors
Lab products found in correlation
Anti ubiquitin, by Merck Group (1 mentions)
Odyssey classic reader, by LI COR (1 mentions)
Odyssey software package 3, by LI COR (1 mentions)
Gs 700 imaging densitometer, by Bio-Rad (1 mentions)
Lc3 1 2, by Novus Biologicals (1 mentions)
Enhanced chemi luminescence (ecl), by Cytiva (1 mentions)
Odyssey infrared imaging system, by LI COR (1 mentions)
β actin, by Merck Group (1 mentions)
Quantity one software, by Bio-Rad (1 mentions)
Tissuelyser, by Qiagen (1 mentions)
Fetal bovine serum (fbs), by Atlanta Biologicals (1 mentions)
2 protocols using gapdh and α tubulin
1
Protein Extraction and Western Blotting
2
Cell Culture Reagents and Antibodies
Check if the same lab product or an alternative is used in the 5 most similar protocols
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All cell culture reagents were purchased from Invitrogen Technologies (Eugene, OR) except fetal bovine serum (FBS), which was obtained from Atlanta Biologicals (Lawrenceville, GA). Unless stated otherwise, all additional chemicals were obtained from Sigma-Aldrich (St Louis, MO). The anti-α-SMA and anti-vimentin antibodies were obtained from Sigma-Aldrich, collagen types I and III and β-arrestin-1 and -2 from Abcam (Cambridge, MA), phospho-p44/42 MAPK and ERK1/2 from Cell Signaling Technology, Inc. (Danvers, MA), and GAPDH and α-tubulin from Santa Cruz Biotechnologies (Santa Cruz, CA).
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