Immunostaining was performed using formalin-fixed paraffin-embedded tumor specimens and a rabbit monoclonal anti-CD20 antibody (1:250; Abcam), as previously described.17 (link)
Anti cd20 antibody
Manufactured by Abcam
Sourced in Japan
Anti-CD20 antibody is a laboratory reagent used for the detection and analysis of the CD20 protein, which is expressed on the surface of B cells. The antibody can be used in various immunological techniques, such as flow cytometry, immunohistochemistry, and Western blotting, to identify and study B cells.
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Lab products found in correlation
Rabbit polyclonal anti cd68 antibody, by Abcam (1 mentions)
Streptavidin peroxidase conjugate, by Zhongshan Biotechnology (1 mentions)
Anti cd3 antibody, by Thermo Fisher Scientific (1 mentions)
3 3 diaminobenzidine (dab), by Nichirei Biosciences (1 mentions)
Histofine simple stain max po multi, by Nichirei Biosciences (1 mentions)
Hematoxylin dye, by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole dapi solution, by Merck Group (1 mentions)
Anti cd3 antibody, by Abcam (1 mentions)
Protein a g plus agarose, by Santa Cruz Biotechnology (1 mentions)
Np 40 lysis buffer, by Beyotime (1 mentions)
5 protocols using anti cd20 antibody
1
Immunostaining of CD20 in Tumors
2
Immunohistochemical Analysis of Immune Cell Markers
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Paraffin-embedded tissues were sectioned at the thickness of 5-μm for immunohistochemistry staining. Sections were deparaffinized and rehydrated, endogenous peroxidases were inactivated with methanol containing 0.3% hydrogen peroxide for 30 min. Antigen retrieval was performed with citrate buffer (pH 6) at 95 °C for 30 min. After incubation in blocking solution (5% normal goat serum) for 10 min at room temperature, the slides were incubated with anti-CD3 antibody (Invitrogen, 1:200), anti-CD20 antibody (Abcam, 1:200) and rabbit anti-CD68 polyclonal antibody (Abcam, 1:200) overnight at 4 °C. After three washes, the sections were incubated with biotinylated anti-IgG at 37 °C for 1 h, followed by Streptavidin-peroxidase conjugate (Zhongshan Biotechnology). Immunoreactivity was detected using 3, 30 diaminobenzidine and the sections were counterstained with hematoxylin for observation by microscopy.
3
Immunohistochemical Analysis of Gastric Tissue
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Tissue sections were deparaffinized in xylene and rehydrated in an ethanol series with phosphate-buffered saline. Antigen activation was performed in an autoclave at 121°C for 20 min with citrate buffer solution. Endogenous peroxidase was blocked with 3% hydrogen peroxide for 10 min. To eliminate non-specific staining, the sections were incubated with 5% normal goat serum for 20 min. Sections were then incubated with anti-hepcidin antibody (diluted 1:1600; Abnova, Taipei City, Taiwan), anti-H + /K + ATPase antibody (diluted 1:6000; BMC, Inc., Tokyo, Japan), and anti-CD20 antibody (diluted 1:100; Abcam plc, Cambridge, UK) at 4°C overnight. After primary antibody treatment, the sections were incubated with Histofine Simple Stain MAX-PO MULTI (Nichirei Bioscience, Tokyo, Japan) for 40 min. After washing with phosphate-buffered saline, the sections were incubated with DAB (Nichirei Bioscience, Tokyo, Japan) and immediately washed with tap water after color development. Finally, the sections were counterstained with Mayer's hematoxylin, dehydrated, and mounted for microscopic observation.
4
Immunohistochemistry Protocol for Cell Markers
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The formaldehyde solution, methanol solution, xylene, anhydrous ethanol, and other reagents were purchased from Guangdong Guanghua Chemical Factory (Guanghua, Guangdong, China). The 5% skimmed milk solution was purchased from Biyuntian Biotechnology Company (Biyuntian Biotechnology, Beijing, China). The rabbit anti-cluster of differentiation (CD) 80 (CD80) antibody was purchased from Abcam (Abcam, Cambridge, UK). The mice anti-PRRSV-N protein-specific antibody was purchased from RTI Corporation (RTI, Anyang-City, Korea). The anti-CD20 antibody (Abcam), anti-CD3 antibody (Abcam), and mouse anti-calgranulin + calprotectin antibody were purchased from Abcam. Alexa Fluor 488-labeled sheep anti-rabbit antibody (Abcam) and Alexa Fluor 647-labeled sheep anti-mouse antibody (Abcam) were purchased from Abcam Company. The 4',6-diamidino--2-phenylindole (DAPI) solution, hematoxylin dye and 0.1% Tween20 solution were purchased from Sigma-Aldrich (Sigma-Aldrich, St. Louis, MO, USA).
5
Immunoprecipitation of CD20 from OCI-ly7 cells
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