Cells were lysed with RIPA Lysis Buffer (P0013B, Beyotime, China) or Nuclear and Cytoplasmic Protein Extraction Kit (P0027, Beyotime, China). Then protein concentrations were quantified with BCA Protein Assay Kit (P0011, Beyotime, China). Twenty microlitre of proteins were separated by SDS-PAGE and then transferred onto PVDF membranes. After blocking, the membranes were incubated with the one of following antibodies: anti-CARMA3 antibody (1:1000, bs-7081R, Bioss, China), anti-MMP2 antibody (bs-0412R, Bioss), anti-MMP9 antibody (1:1000, bs-4593R, Bioss, China), anti-β-catenin antibody (1:1000, #8480, Cell Signaling Technology, USA), anti-Survivin antibody (1:1000, #2808, Cell Signaling Technology, USA), anti-C-myc antibody (1:500, sc-40, Santa Cruz Biotechnology, USA), anti-beta-Actin (1:5000, bsm-33036 M, Bioss, China), or anti-Histone H3.1 (1:500, bs-17422R, Bioss, China) at 4 °C overnight. Then the membranes were incubated in HRP-labeled Goat Anti-Rabbit IgG (H + L) (1:5000, A0208, Beyotime, China) or HRP-labeled Goat Anti-Mouse IgG (H + L) (1:5000, A0216, Beyotime, China) at 37 °C for 45 mins. After incubating ECL reagent for 5 mins, the proteins were detected by ECL Select Western Blotting Detection System (Beyotime, China).
Bs 7081r
Manufactured by Bioss Antibodies
Sourced in China
The Bs-7081R is a lab equipment product manufactured by Bioss Antibodies. It is a device designed for laboratory use, but a detailed factual description of its core function cannot be provided while maintaining an unbiased and concise approach. Further information is required to give a complete and accurate depiction of this product.
Automatically generated - may contain errors
Lab products found in correlation
Bs 4593r, by Bioss Antibodies (1 mentions)
Anti beta actin, by Bioss Antibodies (1 mentions)
Anti c myc antibody, by Santa Cruz Biotechnology (1 mentions)
Hrp labeled goat anti mouse igg h l, by Beyotime (1 mentions)
Bsm 33036m, by Bioss Antibodies (1 mentions)
Anti survivin antibody, by Cell Signaling Technology (1 mentions)
Bs 17422r, by Bioss Antibodies (1 mentions)
Anti β catenin antibody, by Cell Signaling Technology (1 mentions)
Ripa lysis buffer, by Beyotime (1 mentions)
Bca protein assay kit, by Beyotime (1 mentions)
Nuclear and cytoplasmic protein extraction kit, by Beyotime (1 mentions)
Hrp labeled goat anti rabbit igg h l, by Beyotime (1 mentions)
H8070, by Solarbio (1 mentions)
Anti β catenin, by Cell Signaling Technology (1 mentions)
Hrp labeled streptavidin, by Beyotime (1 mentions)
Biotin labeled goat anti rabbit igg h l, by Beyotime (1 mentions)
Hematoxylin, by Solarbio (1 mentions)
3 3 diaminobenzidine (dab), by Solarbio (1 mentions)
Ab238004, by Abcam (1 mentions)
Bs 10090r, by Bioss Antibodies (1 mentions)
3 protocols using bs 7081r
1
Western Blot Analysis of Protein Targets
2
Immunohistochemical Analysis of CARD10 and β-Catenin
Check if the same lab product or an alternative is used in the 5 most similar protocols
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The tumor nodules from mice were subsequently removed and fixed into paraffin sections. The fixed tissue paraffin sections were first incubated in 3% hydrogen peroxide for 15 mins at RT. After blocking, they were incubated with anti-CARD10 (1:200, bs-7081R, Bioss, China) or anti-β-Catenin (1:100, #8480, Cell Signaling Technology, USA) at 4 °C overnight, and then incubated in Biotin-labeled Goat Anti-Rabbit IgG (H + L) (1:200, A0277, Beyotime, China) at 37 °C for 30 mins. Thereafter, the protein signals were magnified with HRP-labeled Streptavidin (1:200, A0303, Beyotime, China), and visualized with DAB (DA1010, Solarbio, China). Cell nuclei were counterstained with hematoxylin (H8070, Solarbio, Beijing, China). Images were captured under a microscope.
3
Immunohistochemical Staining of Tissue Chips
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We performed an immunohistochemical staining assay on the tissue chips in accordance with a previously described method [48 (link)]. The antibodies we used included anti-GPC2 (1:200, AF2304SP, Goat, IgG, Novus), anti-CNOT6L (1:75, abs108959, Rabbit, IgG Absin), anti-FASN (1:300, 66,591-1-Ig, Mouse, IgG, Proteintech), anti- MAP2 (1:2500, 66,846-1-Ig, Rabbit, IgG, Proteintech), anti-BMP6 (1:500, bs-10090R, Rabbit, IgG, Bioss), anti-CARD10 (1:300, bs-7081R, Rabbit, IgG, Bioss), anti-GALNT12 (1:100, ab201196, Rabbit, IgG, Abcam), anti-IgG (ab238004, Mouse, Abcam), anti-IgG (A7007, Goat, Beyotime), and anti-IgG (30,000-0-AP, Rabbit, Proteintech). The IRS (value, 0–12) was calculated by multiplying the staining strength grade by the positive area grade. The grade of staining strength was prescribed below: 0, negative; l, weak; 2, moderate; and 3, strong. The positive area grade was described as follows: zero-grade, less than 5%; first-grade, 5% to 25%; second-grade, 26% to 50%; third-grade, 51% to 75%; and fourth-grade, greater than 75%.
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