Cell lysates were run on 4–12% gradient SDS-PAGE gels under reducing conditions as previously described.29 (link) Western blot was performed using primary antibodies against 15LO1 (1: 1000 dilution, Gift of Dr. Doug Conrad, University of California, San Diego, USA), CCL26 (1:100 dilution, R&D, USA), phospho-ERK (pERK, 1: 5000 dilution, Sigma, USA), total-ERK (tERK, 1: 5000 dilution, Sigma, USA), and GAPDH (1: 1000 dilution, Novus, USA). Densitometry analysis was performed using Image J Software. The densitometry of pERK was presented as a ratio over tERK and the densitometry of 15LO1 and CCL26 were presented as a ratio over GAPDH. For details, see the online supplement .
Ccl26
Manufactured by R&D Systems
Sourced in United States
CCL26 is a recombinant human chemokine that belongs to the CC chemokine family. It functions as a chemoattractant for eosinophils.
Automatically generated - may contain errors
Lab products found in correlation
Gapdh, by Novus Biologicals (2 mentions)
Sds page gel, by Thermo Fisher Scientific (1 mentions)
Slc7a11, by Cell Signaling Technology (1 mentions)
Polyvinylidene difluoride membrane, by Thermo Fisher Scientific (1 mentions)
Amersham imager 600, by GE Healthcare (1 mentions)
Supersignal west femto maximum sensitivity substrate, by Thermo Fisher Scientific (1 mentions)
β actin, by Merck Group (1 mentions)
2 protocols using ccl26
1
SDS-PAGE and Western Blot Analysis
2
Western Blot Analysis of Cellular Proteins
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Cell lysates and basilar supernatants were run in 4%–12% sodium dodecyl sulphate–polyacrylamide (SDS-PAGE) gels (Invitrogen) and transferred onto polyvinylidene difluoride membrane (Invitrogen). For supernatant studies, 40 μL of culture media was loaded as previously described (14 (link)).
Primary antibodies against SLC7A11 (Cell Signaling Technology, 12691, 1:500 dilution), iNOS (BD Biosciences, 610329, 1:500 dilution), 15LO1 (Abnova, H00000246-D01P, 1:1000 dilution), GPX4 (Abcam, 125066, 1:1000 dilution), POSTN (Santa Cruz Biotechnology, 67233, 1:500), and CCL26 (R&D Systems, AF653, 1:500 dilution) were used. GAPDH (Novus Biologicals, NB300-320, 1:1000 dilution) or β-actin (Sigma-Aldrich, A5441, 1:2000 dilution) was measured as control. Membranes were developed using an Amersham Imager 600 (GE Healthcare Life Sciences) with SuperSignal West Femto Maximum Sensitivity Substrate (Thermo Fisher Scientific, 34096). Densitometry analysis was performed using ImageJ software (NIH). See complete unedited blots inSupplemental Figure 5 .
Primary antibodies against SLC7A11 (Cell Signaling Technology, 12691, 1:500 dilution), iNOS (BD Biosciences, 610329, 1:500 dilution), 15LO1 (Abnova, H00000246-D01P, 1:1000 dilution), GPX4 (Abcam, 125066, 1:1000 dilution), POSTN (Santa Cruz Biotechnology, 67233, 1:500), and CCL26 (R&D Systems, AF653, 1:500 dilution) were used. GAPDH (Novus Biologicals, NB300-320, 1:1000 dilution) or β-actin (Sigma-Aldrich, A5441, 1:2000 dilution) was measured as control. Membranes were developed using an Amersham Imager 600 (GE Healthcare Life Sciences) with SuperSignal West Femto Maximum Sensitivity Substrate (Thermo Fisher Scientific, 34096). Densitometry analysis was performed using ImageJ software (NIH). See complete unedited blots in
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