PFC samples collected immediately following decapitation under anesthesia were lysed in ice-cold lysis buffer containing protease inhibitor cocktail (1860932, ThermoFisher scientific, Waltham, MA) and protein concentration was determined by bicinchoninic acid (BCA) assay (MilliporeSigma St. Louis, MO). Protein was electrophoretically separated on a SDS PAGE gel and transferred to a nitrocellulose membrane. Blots were incubated in the appropriate primary antibody specific for C3 (sc-28294, Santa Cruz Biotech, Dallas, Texas), C3aR (sc-133172; Santa Cruz Biotech),VCAM-1 (114441-1-AP; Proteintech, Rosemont, IL), ICAM-1 (10831-1-AP; Proteintech), β-tubulin (2146, Cell Signaling, Danvers, MA) or β-actin (4967, Cell Signaling); and developed with the ECL Plus Western Blotting Detection System (GE Healthcare). Optical densities of the bands were analyzed using ImageJ software (NIH). For analysis, protein levels were normalized to β-tubulin or β-actin levels then expressed as a fold change of that in control animals.
Icam 1 10831 1 ap
Manufactured by Proteintech
ICAM-1 (10831-1-AP) is a primary antibody that detects the Intercellular Adhesion Molecule 1 (ICAM-1) protein. ICAM-1 is a cell surface glycoprotein that plays a role in the adhesion of cells to the extracellular matrix and to other cells.
Automatically generated - may contain errors
Lab products found in correlation
Ecl plus western blotting detection system, by GE Healthcare (1 mentions)
Bca assay, by Merck Group (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
β tubulin, by Cell Signaling Technology (1 mentions)
Protease inhibitor cocktail, by Thermo Fisher Scientific (1 mentions)
Sc 28294, by Santa Cruz Biotechnology (1 mentions)
Sc 133172, by Santa Cruz Biotechnology (1 mentions)
Pannoramic midi, by 3DHISTECH (1 mentions)
Gapdh, by Proteintech (1 mentions)
Pannoramic viewer, by 3DHISTECH (1 mentions)
2 protocols using icam 1 10831 1 ap
1
Western Blot Analysis of Complement and Adhesion Proteins
2
Immunohistochemistry and Western Blot Analysis
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IHC staining was performed as previously described.53 (link) Tissue microarrays were scanned with a digital slide scanner (Pannoramic MIDI, 3D HISTECH) after staining and processed with Pannoramic viewer software. Intensity of staining in cells was automatic calculated by Quant center software. Histochemistry score (H-score) was acquired according to the formula: H-score = (percentage of weak intensity area ×1) + (percentage of moderate intensity area ×2) + (percentage of strong intensity area ×3). Western blot analysis was conducted as previously described.54 (link) The following primary antibodies were used: ICAM1 (10831-1-AP, Proteintech), phospho-IκBα (#2859, CST), NF-κB p65 (#8242, CST), phospho-NF-κB p65 (#3033, CST), ALPK1 (GTX01077, GeneTex) and GAPDH (10494-1-AP, Proteintech).
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