The western blot analysis was carried out as described by Park et al. [18 (link)]. The protein sample (20 μg) was loaded and separated by SDS–PAGE. After electrophoresis, the protein was electrotransferred to a poly vinylidene difluoride membrane (Immobilon-P; Millipore Corp., Billeria, MA). The antibodies phosphorylated Src (Y416, 1:500), total Src (1:1000), p-Rb (S780, 1:500 dilution), total Rb (1:1,000 dilution), p-IkBα (S32/36, 1:500 dilution), total IkBα (1:1,000 dilution), caspase 8 (1:1,000 dilution), and GAPDH (1:1,000 dilution) were purchased from Cell Signaling Technology (Denver, MA). Antibodies for VEGF (1:200 dilution) and TNFα (1:200 dilution) were purchased from Santa Cruz Biotechnology. Antibody for cyclin D1 (1:500 dilution) and TRβ (1:500) were purchased from Neomarkers (Thermo Scientific, Cambridge, MA) and Rockland, respectively. The blots were stripped with Re-Blot Plus (Millpore, Billeria, MA) and reprobed with rabbit polyclonal antibodies to GAPDH. Band intensities were quantified by using NIH IMAGE software (Image J 1.47).
Total ikbα
Manufactured by Cell Signaling Technology
Total IκBα is an ELISA kit that quantifies the total cellular levels of the inhibitory protein IκBα. IκBα is a critical regulator of the NF-κB signaling pathway, which controls the expression of genes involved in immune, inflammatory, and apoptotic responses.
Automatically generated - may contain errors
Lab products found in correlation
Gapdh, by Cell Signaling Technology (2 mentions)
Caspase 8, by Cell Signaling Technology (1 mentions)
Total rb, by Cell Signaling Technology (1 mentions)
Antibodies for vegf, by Santa Cruz Biotechnology (1 mentions)
Total src, by Cell Signaling Technology (1 mentions)
Immobilon p, by Merck Group (1 mentions)
Phospho ikbα, by Cell Signaling Technology (1 mentions)
Cycloheximide (chx), by Research Products International (1 mentions)
Unc3230, by Cayman Chemical (1 mentions)
Y 27632, by Merck Group (1 mentions)
α smooth muscle actin antibody, by Merck Group (1 mentions)
Bay 11 7082, by MedChemExpress (1 mentions)
Sb203580, by MedChemExpress (1 mentions)
2 protocols using total ikbα
1
Western Blot Analysis of Signaling Proteins
2
Integrin Activation and Signaling Protocols
Check if the same lab product or an alternative is used in the 5 most similar protocols
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GAPDH, phospho-Ik Bα, total Ik Bα, and Rho kinase antibodies were purchased from Cell Signaling Technology. PIP5K1γ antibody was purchased from Abcam. α-smooth muscle actin antibody was purchased from Sigma. Integrin antibodies used were monoclonal anti-active integrin β1 (HUTS-4, Millipore), activating integrin β1 (TS2/16, Invitrogen). c15 was purchased from Tocris. β1-CHAMP was synthesized in our laboratory (16 (link)). Manganese was purchased from Fisher. Y-27632 was purchased from Sigma. SB 203580 and BAY 11–7082 were purchased from MedChemExpress. UNC3230 was purchased from Cayman Chemical. PI(4 (link),5 (link))P2 diC16 was purchased from Echelon Biosciences and used with carrier 1 per manufacturer instructions. Cycloheximide was purchased from Research Products International (RPI).
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