The following monoclonal antibodies (mAbs), secondary reagents and isotype controls used in the study were purchesed from BD Biosciences Pharmingen (Mountain View, CA, USA): phycoerythrin (PE)-conjugated anti-CD4 (clone OX-38), peridinin–chlorophyll–protein (PerCP)-conjugated anti-CD90 (Thy-1.1, clone OX-7), fluorescein isothiocyanate (FITC)/PE/biotin/allophycocyanin (APC)-conjugated anti-CD8 (clone OX-8), PerCP-conjugated anti-TCRαβ (clone R73), PE-conjugated anti-CD45RC (clone OX-22), FITC-conjugated anti-CD2 (clone OX-34), FITC/Alexa Fluor® 488-conjugated anti-Ki67 (clone B56), PE-conjugated anti-CD28 (clone JJ319), PerCP-conjugated streptavidin and FITC-conjugated goat anti-mouse Ig Ab. Additionaly, FITC-conjugated anti-Foxp3 (clone FJK-16s), PerCP eFluor® 710-conjugated anti-CD25 (clone OX-39), APC-conjugated anti-CD4 (clone OX-35) (all eBioscience, San Diego, CA, USA), FITC-conjugated anti-granzyme B (clone GB11), and Alexa Fluor® 647-conjugated anti-TCRαβ (clone R73) (both BioLegend, San Diego, CA, USA) mAbs were also used. Unconjugated polyclonal anti-CD69 (P-17) Ab was obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA), FITC-conjugated rabbit anti-goat IgG Ab and merocyanine dye (MC540) from Sigma–Aldrich Chemie GmbH, and unconjugated mouse anti-CDKN2A/p16INK4a (clone 2D9A12) mAb from Abcam (Cambridge, UK).
Fitc conjugated anti granzyme b clone gb11
Manufactured by BioLegend
Sourced in United States
FITC-conjugated anti-granzyme B (clone GB11) is a monoclonal antibody that recognizes the granzyme B protein. Granzyme B is a serine protease involved in the cytotoxic T-cell and natural killer cell-mediated apoptosis pathway. The FITC conjugation allows for fluorescent detection of granzyme B-positive cells.
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2 protocols using fitc conjugated anti granzyme b clone gb11
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Flow Cytometric Characterization of Immune Cell Subsets
2
Multicolor Flow Cytometry Immunophenotyping
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For immunolabeling, the following monoclonal antibodies (mAbs) were used: fluorescein isothiocyanate (FITC)/phycoerythrin (PE)-conjugated anti-CD161a (clone 10/78), FITC/PE/allophycocyanin (APC)-conjugated anti-CD8 (clone OX-8), peridinin-chlorophyll-protein (PerCP)-conjugated anti-TCRαβ (clone R73), PE-conjugated anti-CD4 (clone OX-38), biotin-conjugated anti-CD80 (clone 3H5), FITC-conjugated anti-RT1B (MHC II, clone OX-6), biotin-conjugated anti-CD86 (clone 24F), FITC-conjugated anti-IFN-γ (clone DB-1), and PE-conjugated anti-IL-10 (clone A5-4). These mAbs as well as PerCP-conjugated streptavidin and PE-conjugated donkey anti-rabbit F(ab’)2 IgG as a second step reagents were obtained from BD Biosciences (Mountain View, CA, USA). Following mAbs were also used: PE-conjugated anti-αOX-62 integrin subunit (anti-OX62; clone OX-62) from Serotec (Oxford, UK), PerCP-eFluor® 710-conjugated anti-CD25 (clone OX39) and FITC-conjugated anti-Foxp3 (clone FJK-16s) from eBioscience, FITC-conjugated anti-granzyme B (clone GB11) from BioLegend (San Diego, CA, USA) and APC-conjugated anti-CXCR3 (clone 868013, R&D Systems, Inc., Minneapolis, MN, USA). Rabbit anti-CX3CR1 was purchased from Abcam (Cambridge, UK).
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