Materials were from Sigma Aldrich (St. Louis, Mo, USA) except rK39 immunochromatographic test strips (InBios International, Seattle, WA, USA), anti-human CD4 (clone 4B12, Novocastra, Newcastle upon Tyne, UK,), Chemokine Receptor 4 (CCR4, clone 1G1), CD3 Peridinin chlorophyll (PerCP, clone SK7), CD8 Alexa Fluor 488 (clone RPA-T8), CD8 PerCP (clone SK1), CCR4 Phycoerythrin (PE, clone 1G1, BD Biosciences, San Jose, CA, USA), PD-1(clone EH12.2H7) and CD127 (clone A019D5) APC (Biolegend, San Diego, CA, USA), IL-5 (clone 9906, R&D Systems, Minneapolis, MN, USA), CCL17 (clone N-20), IL-10 (clone E-10), PD-1 (clone C-20), Perforin H (clone H-315), Granzyme B (clone 2C5, Santa Cruz Biotechnology, Dallas, TX, USA), antibody diluent, Target Retrieval Solution Citrate pH 6 (S2369), secondary detection system EnVision™ G|2 System/AP-Rabbit/Mouse (Permanent Red), EnVision™ FLEX Target Retrieval Solution, EnVision™ DuoFLEX Doublestain System (Dako, Glostrup, Denmark), RNALater and RNAqueous kit (Ambion, Austin, TX, USA), One-step reverse transcription kit from Qiagen (Hilden, Germany) and Bio-Plex Pro™ Human Chemokine Panel 40-Plex (BioRad, Hercules, CA, USA).
Envision duoflex doublestain system
Manufactured by Agilent Technologies
Sourced in Denmark
The EnVision™ DuoFLEX Doublestain System is a laboratory equipment product offered by Agilent Technologies. It is designed to enable the simultaneous detection and visualization of two target molecules within a single sample.
Automatically generated - may contain errors
Lab products found in correlation
Cd8 percp clone sk1, by BD (1 mentions)
Envision g 2 system ap rabbit mouse, by Agilent Technologies (1 mentions)
One step reverse transcription kit, by Qiagen (1 mentions)
Bio plex pro human chemokine panel 40 plex, by Bio-Rad (1 mentions)
Envision flex target retrieval solution, by Agilent Technologies (1 mentions)
2 protocols using envision duoflex doublestain system
1
Multiplex Analysis of Immune Markers
2
Quantification of Cytokines and Immune Cells in Lung
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Quantification of Cytokines and Growth Factors in Plasma by ELISA. Protein levels of TNF-a, VEGF, IL-2, IL-10, IFN-g, TGF-b, and IL-4 were quantified in blood (plasma) from all patients by using ELISA kits (Gen-probe Diaclone SAS, Besançon, France) following previous studies. 15, 35, 36 Quantification of Cytokines and Growth Factors in Lung Tissue (Tumor and Nontumor) by ELISA. Protein levels of TNF-a, VEGF, IL-2, IL-10, IFN-g, EGFR, TGF-b, and IL-4 were quantified in lung specimens from all patients with ELISA kits (Raybiotech Inc., Norcross, GA, and Cloud-Clone Corp, Houston, TX), following previous studies. 15, 35, 36 Counts and Types of Macrophages in Lung Specimens (Tumor and Nontumor). M1 and M2 macrophages were identified on 3-mm paraffin-embedded lung sections by using double-staining immunohistochemical procedures (Envision DuoFLEX Doublestain System, Dako North America, Inc., Carpinteria, CA) following the manufacturer's instructions and previous studies. [37] [38] [39]
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