Antibodies were obtained from the following suppliers: anti-human CD3 (BD Biosciences, 555335), anti-human CD8 (BD Biosciences 555366), anti-human CD107a (BD Biosciences 555801), anti-human CD137 (BD Biosciences 555956). Cell surface expression of ErbB2 was detected by biotylated anti-ErbB2 Affibody (Abcam, ab31890), and EGFR by FITC conjugated anti-EGFR affibody (Abcam, ab81872). EGFR, ErbB2 and CD19 specific CAR expression were detected by biotin-labeled polyclonal anti-human F(ab)2 antibody for (EGFR CAR) or anti-mouse F(ab)2 antibody (for ErbB2 and CD19 CARs)(Jackson Immunoresearch). Samples were then stained with PE-conjugated anti-human IgG Fc Ab (eBioscience, 12-4998-82) or phycoerythrin-labeled streptavidin (eBioscience, 17-4317-82). Flow cytometry acquisition was performed on either a BD FacsCalibur or Accuri C6 Cytometer (BD Biosciences). Analysis was performed using FlowJo software (Treestar).
Anti human cd107a
Manufactured by BD
Sourced in United States
Anti-human CD107a is a laboratory reagent used for the detection and analysis of CD107a expression on the surface of cells. CD107a, also known as LAMP-1, is a lysosome-associated membrane protein that is transiently expressed on the cell surface during degranulation, a process associated with the activation of certain immune cells. This reagent can be used in flow cytometry and other immunological assays to measure the degree of cell activation or degranulation.
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Lab products found in correlation
Anti human cd3, by BD (3 mentions)
Anti human cd8, by BD (3 mentions)
Anti human cd137, by BD (2 mentions)
Phycoerythrin labeled streptavidin, by Thermo Fisher Scientific (2 mentions)
Facscalibur, by BD (2 mentions)
Accuri c6 cytometer, by BD (2 mentions)
Anti mouse f ab 2 antibody, by Jackson ImmunoResearch (2 mentions)
Ab81872, by Abcam (1 mentions)
Brefeldin a, by Merck Group (1 mentions)
Monensin, by BD (1 mentions)
Rosettesep nk cell enrichment kit, by STEMCELL (1 mentions)
Ique analyzer, by Intellicyt (1 mentions)
Maxisporp elisa plates, by Thermo Fisher Scientific (1 mentions)
Rhil 15, by STEMCELL (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Fix perm cell permeabilization kit, by Thermo Fisher Scientific (1 mentions)
Golgiplug, by BD (1 mentions)
Anti human cd86, by BD (1 mentions)
Anti human tim 3, by BD (1 mentions)
Anti human pd 1, by BD (1 mentions)
5 protocols using anti human cd107a
1
Multiparametric Flow Cytometry Characterization
2
T Cell Surface Marker Detection
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Antibodies were obtained as follows: anti-human CD3 (BD Biosciences, 555335), anti-human CD8 (BD Biosciences, 555366), anti-human CD107a (BD Biosciences, 555801), and anti-human CD137 (BD Biosciences, 555956). The antibodies were incubated with T cells at 4°C for 25 min and washed twice (PBS with 2% FBS). Mesothelin CAR, ErbB2 CAR, CD19 CAR, and OKT3-28BB expression were detected by biotin-labeled polyclonal anti-mouse F(ab)2 antibody (Jackson Immunoresearch). Samples were then stained with phycoerythrin-labeled streptavidin (eBioscience, 17-4317-82). Flow cytometry acquisition was performed on either a BD FACSCalibur or Accuri C6 Cytometer (BD Biosciences). Analysis was performed using FlowJo software (Treestar).
3
Antibody-dependent NK cell activation
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To determine Antibody-dependent NK cell activation, MaxiSporp ELISA plates (Thermo Fisher) were coated with respective antigen for 2h at RT and then blocked with 5% BSA (Sigma-Aldrich). 50 μl 1:50 diluted plasma sample or monoclonal Abs was added to the wells and incubated overnight at 4°C. NK cells were isolated from buffy coats from healthy donors using the RosetteSep NK cell enrichment kit (STEMCELL Technologies, MA, USA) and stimulated with rhIL-15 (1ng/ml, STEMCELL Technologies) at 37°C overnight. NK cells were added to the washed ELISA plate and incubated together with anti-human CD107a (BD, 1:40, clone: H4A3), brefeldin A (Sigma-Aldrich, MO, USA), and monensin (BD) for 5 hours at 37°C. Next, cells were surface stained for CD56 (BD, 1:200, clone: B159), CD16 (BD, 1:200, clone: 3G8), and CD3 (BD, 1:800, UCHT1). After fixation and permeabilization with FIX & PERM Cell Permeabilization Kit (Thermo Fisher), cells were stained for intracellular markers MIP1β (BD, 1:50, clone: D21–1351) and IFNγ (BD, 1:17, clone: B27). NK cells were defined as CD3-CD16+CD56+ and frequencies of degranulated (CD107a+), INFγ+ and MIP1β+ NK cells determined on an iQue analyzer (Intellicyt)(49 (link)).
4
NK Cell Cytotoxicity Assay
5
Cytokine and phenotypic analysis of mature DCs co-cultured with ASALs
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Mature DCs were cocultured with ASALs at a 1:4 ratio in 96-well round bottom plates in 200 µl of culture media. Enzyme-linked immunosorbent assay kits (Mabtech, Nacka Strand, Sweden) were used to measure IFN-γ, IL-2, and IL-12p70 in the supernatants. Phenotypic analysis of T cells was performed using the following fluorophore-labeled mouse monoclonal antibodies: anti-human CD3, anti-human CD4, anti-human CD8, anti-human CD28, anti-human CD27, anti-human CD107a, anti-human Tim3, anti-human PD1, anti-human CD3 zeta chain, and matched mouse IgG isotype controls (all from BD Biosciences). Phenotypic analysis of DCs was performed using the following fluorophore-labeled mouse monoclonal antibodies: anti-human CD80, anti-human CD86, anti-human CD70, and anti-human CD64 expression and matched mouse IgG isotype controls (all from BD Biosciences). These data were on analysis BD FACS Canto II (BD Biosciences).
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