Alexa Fluor 700-labeled anti-CD3, BUV496-labeled anti-CD4, allophycocyanin H7-labeled anti-CD45RA, and BUV395-labeled anti-Ki-67 were obtained from BD Biosciences. BV785-labeled anti-CD8, BV650-labeled anti-CD127, and Pacific Blue-labeled anti-Perforin came from BioLegend. Foxp3/Transcription Factor Staining Buffer Set was from eBioscience. HLA-A*1101 HIV-1 NEF (GAFDLSFFLK) soluble biotinylated monomers were produced at Chulalongkorn University (Bangkok, Thailand) as previously described33 and tetramerized with PE-conjugated extravidin (Sigma-Aldrich). LIVE/DEAD Fixable Aqua Dead Cell Stain Kit (LIVE/DEAD) was from Thermo Fisher Scientific (Molecular Probes).
Pe conjugated extravidin
Manufactured by Merck Group
PE-conjugated extravidin is a laboratory reagent composed of extravidin, a protein derived from egg white, which is conjugated to the fluorescent dye phycoerythrin (PE). This product is commonly used in various immunoassay and cell-based applications requiring the detection and visualization of target molecules.
Automatically generated - may contain errors
Lab products found in correlation
Alexa fluor 700 labeled anti cd3, by BD (2 mentions)
Buv496 labeled anti cd4, by BD (2 mentions)
Allophycocyanin h7 labeled anti cd45ra, by BD (2 mentions)
Buv395 labeled anti ki 67, by BD (2 mentions)
Bv785 labeled anti cd8, by BioLegend (2 mentions)
Bv650 labeled anti cd127, by BioLegend (2 mentions)
Pacific blue labeled anti perforin, by BioLegend (2 mentions)
Live dead fixable aqua dead cell stain kit live dead, by Thermo Fisher Scientific (2 mentions)
Foxp3 transcription factor staining buffer set, by Thermo Fisher Scientific (2 mentions)
L arginine hcl, by Merck Group (1 mentions)
Tris hcl ph 8, by Merck Group (1 mentions)
Oxidized glutathione, by Merck Group (1 mentions)
Na edta, by Merck Group (1 mentions)
Reduced glutathione, by Merck Group (1 mentions)
3 protocols using pe conjugated extravidin
1
Multiparameter Flow Cytometry Analysis
2
Synthesis of MHC-Peptide Tetrameric Complexes
Check if the same lab product or an alternative is used in the 5 most similar protocols
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The synthesis of MHC-peptide tetrameric complexes has been reported previously.59 (link) Briefly, the MHC HLA-A*0201 heavy chain, β2m and epitope peptide (DEPDC1#5 or Melan-A26–35*A27L peptides) were subjected to a refolding in vitro in Tris–HCl pH 8, L -Arginine-HCl, NaEDTA, oxidized glutathione and reduced glutathione (Sigma-Aldrich). The complex was isolated through dialysis, concentrated and purified by HPLC to separate monomers from unconjugated components. Monomers were then enzymatically biotinylated by the enzyme BirA. The biomonomers obtained were purified and then quantified with a spectrophotometer. The tetramers were finally assembled with PE-conjugated extravidin (Sigma-Aldrich) for subsequent use in flow cytometry analysis.
3
Multiparameter Flow Cytometry Analysis
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