PBMCs were labeled with carboxyfluorescein diacetate succinimidyl ester (CFSE; Invitrogen). PBMCs were plated at 107 cells/mL in 500µL into 24-well fibronectin coated plates (Corning Incorporated). Where indicated, cells were incubated with 1µM ibrutinib, 500nM firategrast (VLA-4 antagonist; MedChem Expression), 50ng/mL Phorbol 12-myristate 13-acetate (PMA; Sigma), 10µg/mL αCD18 (Abcam Inc.), 10µg/mL αCD29 (R&D Systems) or 10µg/mL αCD49d (LifeSpan BioSciences, Inc.) for 1 hour at 37°C. The plates were centrifuged at 400×g for one minute and cells were left to adhere for 1 hour at 37°C. Non-adhered cells were removed by washing three times with PBS (Lonza). Wells were imaged using the EVOS fl microscope (Advanced Microscopy Group). Quantification of the number of adhered CLL PBMCs was done using the Image J software (NIH). Experiments were performed in duplicate plates with 4–6 images collected per plate; results are reported as the median cell count per visual field.
αcd29
Manufactured by R&D Systems
αCD29 is a monoclonal antibody that recognizes the human CD29 (Integrin beta 1) protein. CD29 is a cell surface receptor involved in cell-cell and cell-matrix interactions. The αCD29 antibody can be used for the identification and isolation of CD29-positive cells in various applications.
Automatically generated - may contain errors
Lab products found in correlation
Carboxyfluorescein succinimidyl ester (cfse), by Thermo Fisher Scientific (1 mentions)
Phorbol 12 myristate 13 acetate pma, by Merck Group (1 mentions)
Phosphate buffered saline (pbs), by Lonza (1 mentions)
α cd11b, by Thermo Fisher Scientific (1 mentions)
αcd11c, by BioLegend (1 mentions)
Mopc 21, by BioLegend (1 mentions)
Clone mab11, by BioLegend (1 mentions)
2 protocols using αcd29
1
PBMCs Adhesion Assay Protocol
2
Blocking of Fc Receptors in Immunoassays
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The chimeric α-ganglioside GM2 monoclonal antibody (KM966) was a kind gift from Kyowa Hakko Kirin Co., Ltd. The chimeric α-CD20 (rituximab) and the humanized α-HER2 (trastuzumab) were purchased from Roche Pharmaceuticals. The TCR-like antibodies were generated in laboratory of Antonio Bertoletti. α-CD64 (clone 10.1, BioLegend), α-CD32 (clone AT10, Abcam), α-CD32a (clone IV.3, StemCell Technologies), α-CD32b (polyclonal, Abcam), α-CD16 (clone DJ130c, AbD Serotec), α-CD18 (Clone TS1/18, Biolegend), α-CD11a (Clone TS1/22, Thermo Scientific), α-CD11b (Clone CBRM1/5, eBioscience), α-CD11c (Clone 3.9, Biolegend), α-CD29 (Clone P5D2, R&D systems), α-TNFα (Clone Mab11, Biolegend), α-TNFR (Clone mab625, R&D systems) and an isotype-matched control antibody (MOPC21, BioLegend) were used for blocking experiments.
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