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Vcam 1 fc

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VCAM-1-Fc is a recombinant human fusion protein consisting of the extracellular domain of Vascular Cell Adhesion Molecule-1 (VCAM-1) and the Fc region of human IgG1. VCAM-1 is a cell surface sialoglycoprotein involved in cell-cell adhesion.

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Lab products found in correlation

Phosphate buffered saline (pbs), by Thermo Fisher Scientific (2 mentions) Ionomycin, by BioLegend (1 mentions) Foxp3 transcription factor fixation permeabilization kit, by Thermo Fisher Scientific (1 mentions) Piroxicam, by Merck Group (1 mentions) Carboxyfluorescein succinimidyl ester (cfse), by Thermo Fisher Scientific (1 mentions) Mouse cd4 t cell isolation kit, by BioLegend (1 mentions) Mojosort mouse cd3 t cell isolation kit, by BioLegend (1 mentions) Recombinant mouse icam 1 fc, by R&D Systems (1 mentions) Efluor 670, by Thermo Fisher Scientific (1 mentions) Brefeldin a, by BioLegend (1 mentions) Phorbol myristate acetate (pma), by Merck Group (1 mentions) Dnase 1, by Roche (1 mentions) Monensin, by BioLegend (1 mentions) Liberase tl research grade, by Roche (1 mentions) Fitc conjugated rabbit anti mouse igg, by Jackson ImmunoResearch (1 mentions) μ slide vi0, by Ibidi (1 mentions) Human icam 1 fc, by R&D Systems (1 mentions)

5 protocols using vcam 1 fc

1

Detection of High-Affinity β1 by Flow Cytometry

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For detection of high-affinity β1 by flow cytometry, cells were stimulated for different times with CXCL12 and/or VCAM-1-Fc (R&D Systems) and fixed with 2% PFA before adding the HUTS-21 anti-β1 mAb (10 μg/ml) for 30 min at 4 °C. After washing, cells were incubated with Alexa Fluor 488-conjugated rabbit anti-mouse IgG (Jackson ImmunoResearch Laboratories). Fluorescence intensity data indicate fold-induction values relative to those from control untreated cells, which were given an arbitrary value of 1.
Sosa-Costa A., Isern de Val S., Sevilla-Movilla S., Borgman K.J., Manzo C., Teixidó J, & Garcia-Parajo M.F. (2016). Lateral Mobility and Nanoscale Spatial Arrangement of Chemokine-activated α4β1 Integrins on T Cells. The Journal of Biological Chemistry, 291(40), 21053-21062.
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2

Multiparametric Phenotyping of T Cells

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The following antibodies were from BioLegend: CD3 (17A2, 2C11), CD4 (GK1.5), CD8 (53–6.7), CD44 (1M7), CD62L (MEL-14), B220 (RA3-6B2), CD29 (HMβ1-1), CD18 (M18/2), β7 (FIB504), Foxp3 (MF-14), CD28 (37.51), IL-10 (JES5-16E3), and TGF-β1 (TW7-16B4). Secondary Alexa Fluor–labeled antibodies were from Jackson ImmunoResearch. Foxp3 transcription factor fixation/permeabilization kit was purchased from eBioscience. CFSE and eFluor 670 were purchased from Invitrogen and BioLegend respectively. PMA and piroxicam were from Sigma. Ionomycin, brefeldin A, and monensin were from BioLegend. MojoSort mouse CD3 T cell isolation kit and mouse CD4 T cell isolation kit were from BioLegend. Liberase TL (Research Grade) and DNase I were from Roche. Recombinant mouse ICAM-1-Fc and VCAM-1-Fc were from R&D Systems. Recombinant mouse MAdCAM-1-Fc was purified by ProteinA beads as previously described (Sun et al., 2011 (link)).
Sun H., Lagarrigue F., Wang H., Fan Z., Lopez-Ramirez M.A., Chang J.T, & Ginsberg M.H. (2020). Distinct integrin activation pathways for effector and regulatory T cell trafficking and function. The Journal of Experimental Medicine, 218(2), e20201524.
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3

VCAM-1-Fc Cell Binding Assay

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For cell binding to VCAM-1-Fc (R&D Systems), cells were stimulated for 20 s with CXCL12 before VCAM-1-Fc was added, which was detected by flow cytometry using phycoerythrin-conjugated AffiniPure F(ab′)2 fragment goat anti–human immunoglobulin G (IgG), Fcγ-fragment specific (Jackson ImmunoResearch, West Grove, PA). For the detection of high-affinity β1, cells were stimulated for 20 s with CXCL12 before HUTS-21 anti–β1 mAb (10 μg/ml) was added for 30 min at 4ºC. After washing, cells were incubated with FITC-conjugated rabbit anti–mouse IgG (Jackson Immunoresearch). Fluorescence intensity data indicate fold-induction values relative to those from control untreated cells, which were given an arbitrary value of one.
Dios-Esponera A., Isern de Val S., Sevilla-Movilla S., García-Verdugo R., García-Bernal D., Arellano-Sánchez N., Cabañas C, & Teixidó J. (2015). Positive and negative regulation by SLP-76/ADAP and Pyk2 of chemokine-stimulated T-lymphocyte adhesion mediated by integrin α4β1. Molecular Biology of the Cell, 26(18), 3215-3228.
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4

Coating Ibidi Microslides for Cell Adhesion

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Channels Ibidi μ-Slide VI0.1 (Ibidi GMBH, Martinsried, Germany) were coated at 4°C with 50 μL of a 10-μg/mL human ICAM-1-Fc or VCAM-1-Fc (R&D Systems, Minneapolis, MN, United States) in phosphate-buffered saline (PBS) (Gibco), rinsed three times with PBS, then blocked with 75 μL of a 2% bovine serum albumin (BSA, Sigma–Aldrich) solution in PBS (Life Technologies) for 25 min, and rinsed again three times with PBS, and finally filled with RPMI before injection of cells.
Robert P., Biarnes-Pelicot M., Garcia-Seyda N., Hatoum P., Touchard D., Brustlein S., Nicolas P., Malissen B., Valignat M.P, & Theodoly O. (2021). Functional Mapping of Adhesiveness on Live Cells Reveals How Guidance Phenotypes Can Emerge From Complex Spatiotemporal Integrin Regulation. Frontiers in Bioengineering and Biotechnology, 9, 625366.
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5

Generating Pro-T Cells from Human CD34+ Cells

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Human CD34+ cells isolated from PI mouse bone marrow were cultured in Delta-like 4-Fc- (Sino Biological, Beijing, China) and VCAM-1-Fc- (R&D Systems, Minneapolis, MN) coated 96-well plates to generate pro-T cells, as described (43 (link)). Human fetal thymus at gestational age 19 weeks was obtained from Advanced Bioscience Resources, cryopreserved and recovered. 2mm3 fetal thymus fragments were incubated for 12 hours with 100mM 2-deoxyglucose (Carbosynth. Compton, England) to deplete thymocytes. Pro-T cells and thymus fragments were then cocultured as described, for 48 hours as hanging drops, then for 12 days on media-soaked gelfoam sponges (Pfizer, New York, NY) before flow cytometric (FCM) analysis (44 ).
Nauman G., Danzl N.M., Lee J., Borsotti C., Madley R., Fu J., Hölzl M.A., Dahmani A., Gonzalez A.D., Chavez É., Campbell S.R., Yang S., Satwani P., Liu K, & Sykes M. (2022). Defects in long-term APC repopulation ability of adult human bone marrow HSCs compared to fetal liver HSCs. Journal of immunology (Baltimore, Md. : 1950), 208(7), 1652-1663.
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