Cancer tissues were isolated from p16INK4A‐positive and negative CRC. Collected cancer tissues were washed with PBS three times and incubated at 37 °C for 24 h with GFP tagged primary isolated CD8+ T cells in completed RPMI medium. In the case of CXCL12 neutralizing antibody treatment, 5 µg mL−1 of anti‐CXCL12 antibody (R&D System) was applied to the medium for 24 h. The tissues were fixed with formalin for 24 h and perform immunohistochemistry with p16INK4A, CD8 and GFP antibodies.
Anti cxcl12 antibody
Manufactured by R&D Systems
Sourced in United States
The Anti-CXCL12 antibody is a laboratory tool designed for the detection and quantification of CXCL12, a chemokine involved in various biological processes. This antibody can be used in applications such as ELISA, Western blotting, and immunohistochemistry to study the expression and localization of CXCL12 in samples.
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Lab products found in correlation
4 protocols using anti cxcl12 antibody
1
Investigating p16INK4A in CRC Immune Response
2
CXCL12, IL-1α, and IL-1Ra Assays
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Recombinant human CXCL12 and anti-CXCL12 antibody were purchased from R&D systems (Minneapolis, MN), recombinant human IL-1α was provided by Diaclone (Beasancon, France), while recombinant human IL-1 Receptor Antagonist (IL-1Ra) was provided from Pepro Tech EC Ltd (London, UK).
3
Immunohistochemical Scoring of CXCL12 and BRAF
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Immunohistochemical staining was performed on 4 μm-thick sections from TMA blocks. Tissues were stained with anti-CXCL12 antibody (R&D Systems, Minneapolis, MN, USA; 1:50 dilution) and anti-BRAF V600E monoclonal antibody (Spring Bioscience, Pleasanton, CA, USA; 1:50 dilution) using an automated immunostainer (Benchmark XT; Ventana Medical Systems Inc., Tucson, AZ, USA). For immunocytochemical staining of FNA samples, cells on LBC slides were fixed in 95% ethyl alcohol for 30 min and then processed as for immunohistochemistry.
The immunohistochemical and immunocytochemical staining results of TMA and FNA slides were examined independently. The extent of cytoplasmic staining was graded from 0 to 3+ as follows: 0, negative; 1+, focal weak staining; 2+, diffuse weak or focal strong staining; and 3+, diffuse strong staining. Scores of 1+, 2+, and 3+ were considered positive, and a score of 0 was considered negative. All specimens were scored twice by a pathologist (YHM) who was blinded to the mutation analysis results, and the evaluation was repeated by an independent observer. If discrepancies occurred, a consensus score was reached.
The immunohistochemical and immunocytochemical staining results of TMA and FNA slides were examined independently. The extent of cytoplasmic staining was graded from 0 to 3+ as follows: 0, negative; 1+, focal weak staining; 2+, diffuse weak or focal strong staining; and 3+, diffuse strong staining. Scores of 1+, 2+, and 3+ were considered positive, and a score of 0 was considered negative. All specimens were scored twice by a pathologist (YHM) who was blinded to the mutation analysis results, and the evaluation was repeated by an independent observer. If discrepancies occurred, a consensus score was reached.
4
Immunohistochemical Analysis of TROP-2, SLP-2, and CXCL12
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The staining was performed on 4-micrometer thickness tissue samples. Absolute ethyl alcohol was used to remove the paraffin-xylene solution. Tissue sections were incubated for an hour with an anti-TROP-2 antibody (Santa Cruz Biotechnology, USA) at 1:40 dilution, an anti-SLP-2 antibody (Proteintech Group, Chicago, IL, USA) at 1:200 dilution, and an anti-CXCL12 antibody (R&D Systems, Minneapolis, MN, USA) at 1:50 dilution. Mayer’s hematoxylin was used as a counterstain.
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