The cells were lysed in NP-40 lysis buffer containing a protease inhibitor cocktail and a phosphatase inhibitor cocktail (Roche, Rotkreuz, Switzerland). After separation using SDS–PAGE, the proteins were transferred onto nitrocellulose membranes (Bio-Rad, Hercules, USA). Subsequently, the membranes were blocked with 5% non-fat milk in Tris-buffered saline (TBS) containing 0.1% Tween-20 for 1 hour at room temperature. The blots were probed with the relevant primary antibodies overnight at 4°C, washed, and probed with a species-specific horseradish peroxidase-conjugated secondary antibody (Cell Signaling, Beverly, USA). An enhanced chemiluminescence detection method (Pierce ECL Western Blotting Substrate, Thermol, Beverly, MA, USA) was used to visualize the blots. The primary antibodies used were anti-EMMPRIN, anti-MMP-2, anti-uPA, anti-Cathepsin B (Santa Cruz Biotechnology, USA), and anti-β-actin (Sigma–Aldrich, MO, USA).
Anti emmprin
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Anti-EMMPRIN is a laboratory research reagent used to detect and quantify the expression of EMMPRIN (Extracellular Matrix Metalloproteinase Inducer), also known as CD147 or Basigin, in various biological samples. EMMPRIN is a cell surface glycoprotein involved in the regulation of matrix metalloproteinases and cellular processes. This reagent can be utilized in techniques such as Western blot, ELISA, and flow cytometry to study the expression and function of EMMPRIN in different experimental systems.
Automatically generated - may contain errors
Lab products found in correlation
Anti upa, by Santa Cruz Biotechnology (1 mentions)
Anti mmp2, by Santa Cruz Biotechnology (1 mentions)
Species specific horseradish peroxidase conjugated secondary antibody, by Cell Signaling Technology (1 mentions)
Anti cathepsin b, by Santa Cruz Biotechnology (1 mentions)
Anti β actin, by Merck Group (1 mentions)
Nitrocellulose membrane, by Bio-Rad (1 mentions)
Protease inhibitor cocktail, by Roche (1 mentions)
Phosphatase inhibitor cocktail, by Roche (1 mentions)
2 protocols using anti emmprin
1
Western Blot Analysis of Proteolytic Enzymes
2
Immunohistochemical Evaluation of EMMPRIN in Gastric Cancer
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We performed immunostaining with anti EMMPRIN (1:500; Santa Cruz Biotechnology) to confirm EMMPRIN expressions in cultured PMCs and cancer cells.
Then, immunohistochemical staining was analyzed using 110 patients with GCs.
Briefly, slides were deparaffinized and rehydrated with xylene and graded alcohol series and activated by heating. Endogenous peroxidase was blocked and then sections were incubated with the EMMPRIN antibody (1:100; Santa Cruz Biotechnology) for 60 minutes at room temperature, and were incubated with biotinylated rabbit anti-mouse IgG for 10 min. The slides were treated with streptavidin-peroxidase reagent, followed by counterstaining with Mayer's hematoxylin. EMMPRIN expressions in PMCs and cancer cells were evaluated by intensity of stain in peritoneum such as greater omentum, lesser omentum, retroperitoneum and cancer cells of surgical specimen.
Intensity was given scores 0-2 (0; no staining, 1+; weak staining, 2+; strong staining).
The evaluation was performed by two double-blinded distinct observers without being aware of clinical data and outcome.
Then, immunohistochemical staining was analyzed using 110 patients with GCs.
Briefly, slides were deparaffinized and rehydrated with xylene and graded alcohol series and activated by heating. Endogenous peroxidase was blocked and then sections were incubated with the EMMPRIN antibody (1:100; Santa Cruz Biotechnology) for 60 minutes at room temperature, and were incubated with biotinylated rabbit anti-mouse IgG for 10 min. The slides were treated with streptavidin-peroxidase reagent, followed by counterstaining with Mayer's hematoxylin. EMMPRIN expressions in PMCs and cancer cells were evaluated by intensity of stain in peritoneum such as greater omentum, lesser omentum, retroperitoneum and cancer cells of surgical specimen.
Intensity was given scores 0-2 (0; no staining, 1+; weak staining, 2+; strong staining).
The evaluation was performed by two double-blinded distinct observers without being aware of clinical data and outcome.
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