After perfusion with 1% paraformaldehyde, lungs were excised and inflated with agarose at 38°C, further fixed by immersion in 1% paraformaldehyde for 1 hour, washed for 3 hours in PBS, placed in sucrose overnight, embedded in OCT, then sliced into 50-μm sections. These sections were washed with 0.3% Triton X-100 in PBS, incubated for 1 hour in 0.3% Triton X-100/0.2% BSA/0.1% Na+ azide containing 10% donkey serum, then incubated overnight in 0.3% Triton X-100/0.2% BSA/0.1% Na+ azide containing anti-rat CD31 (Pierce; 1:500) and goat anti-c-Met (R&D Systems; 1:500). Sections then were incubated for 6 hours with donkey anti-rat IgG-Alexa Fluor-488 (Life Technologies; 1:500) and donkey anti-goat-IgG-Cy3 (Abcam; 1:500). Washed sections then were fixed in 1% paraformaldehyde in PBS, washed with PBS, and mounted in Vectashield medium with 4',6-diamidino-2-phenylindole (Vector Laboratories, Burlingame, CA).
Goat anti c met
Manufactured by R&D Systems
Goat anti-c-Met is a laboratory reagent used for the detection and analysis of the c-Met protein in biological samples. It is a polyclonal antibody raised in goats against the c-Met protein, which is a receptor tyrosine kinase involved in various cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Rabbit anti phospho cmet, by R&D Systems (2 mentions)
Non immune mouse igg, by Merck Group (2 mentions)
Heated citrate buffer, by Fortis Life Sciences (2 mentions)
Mouse anti matriptase, by R&D Systems (2 mentions)
Rabbit anti phospho cmet, by Abcam (2 mentions)
Rabbit anti c met, by Leica (2 mentions)
Donkey anti goat igg cy3, by Abcam (1 mentions)
3 protocols using goat anti c met
1
Lung Tissue Immunofluorescence Staining
2
Immunohistochemical Analysis of Breast Cancer
Check if the same lab product or an alternative is used in the 5 most similar protocols
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The use of human tissue paraffin embedded samples was approved according to the Institutional Review Board Administration. The “BC20812 and “BRC711” breast cancer tissue arrays with pathology grading were obtained from US Biomax, Inc. (Rockville, MD). Grade II and III tumors were analyzed in this study. Tissue arrays were deparaffinized with xylene and hydrated with graded ethanol solutions. Antigen retrieval was performed using heated citrate buffer, reduced pH (Bethyl Laboratories, Montgomery, TX). The arrays were blocked with 2 % bovine serum albumin in PBS, and immunostained overnight at 4°C5 (link). Primary antibodies were mouse anti-matriptase (1:200), goat anti-c-Met (1:200) and rabbit anti-phospho-cMet (1:200) (R&D Systems, Minneapolis, MN), rabbit anti-matriptase (1:100) (Calbiochem/EMD Millipore, San Diego, CA), rabbit anti c-Met (1:30)(Leica Microsystems Inc., Buffalo Grove, IL) and rabbit anti-phospho-cMet (1:200) (Abcam, Cambridge, MA). As negative controls, non-immune mouse IgG, rabbit IgG or goat IgG were used (Sigma, St. Louis, MO) (adjusted to same final concentration as specific primary antibodies).
3
Immunohistochemical Analysis of Breast Cancer
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