Anti vcam 1

Manufactured by Thermo Fisher Scientific

Sourced in Belgium

Anti-VCAM-1 is a laboratory product that binds to Vascular Cell Adhesion Molecule-1 (VCAM-1). VCAM-1 is a cell surface protein involved in cell-cell adhesion processes. The core function of Anti-VCAM-1 is to detect and quantify VCAM-1 expression.

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Lab products found in correlation

Anti icam 1, by Thermo Fisher Scientific (3 mentions) Cycloheximide (chx), by Santa Cruz Biotechnology (1 mentions) Anti vegfr 1, by Santa Cruz Biotechnology (1 mentions) Anti fgfr 1 antibody, by Santa Cruz Biotechnology (1 mentions) Anti cd31 antibody, by Thermo Fisher Scientific (1 mentions) Anti mcp 1, by Thermo Fisher Scientific (1 mentions) Anti β actin antibody, by Merck Group (1 mentions) Anti vegfr2 antibody, by Cell Signaling Technology (1 mentions) Anti tie2 antibody, by R&D Systems (1 mentions) Anti phospho akt antibody, by Cell Signaling Technology (1 mentions) Anti cd31, by Thermo Fisher Scientific (1 mentions) Anti akt antibody, by Cell Signaling Technology (1 mentions) Recombinant human vegf, by Thermo Fisher Scientific (1 mentions) Anti cd31, by BD (1 mentions) Hepes, by Merck Group (1 mentions) Sodium bicarbonate, by Merck Group (1 mentions) Sorafenib, by LC Laboratories (1 mentions) Sunitinib, by LC Laboratories (1 mentions) Hmvec, by Lonza (1 mentions) Anti tnfα antibodies, by Thermo Fisher Scientific (1 mentions)

5 protocols using anti vcam 1

Evaluation of Anti-Angiogenic Drug Effects

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Sunitinib and sorafenib were from LC Laboratories (Woburn, MA, USA). Sodium bicarbonate and HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid) were from Sigma-Aldrich. Recombinant human VEGF was from Peprotech (#100-20-10). Cycloheximide was purchased from Santa Cruz biotechnology (#SC-3508). For Western blot analysis, the following primary antibodies and concentrations were used: anti-phospho-Akt antibody (1:500) (#4060; Cell Signaling Technology), anti-Akt antibody (1:1000) (#2920; Cell Signaling Technology), anti-VEGFR-2 antibody (1:1000) (#2479; Cell Signaling Technology), anti-VEGFR-1 (1:500) (#sc-316; Santa Cruz biotechnology), anti-β-actin antibody (1:5000) (#A2228; Sigma Aldrich), anti-Tie-2 antibody (1:500) (#MAB313; R&D systems) and anti-FGFR-1 antibody (1:1000) (#sc-121; Santa Cruz Biotechnology). Immunohistochemical staining were performed with anti-CD31 antibody (#Rb-10333-PO; Thermo Scientific). For immunofluorescence anti-CD31 (1:50) (# 553370; BD Pharmigen) and anti-VEGFR-2 antibody (1:50) (#2479; Cell Signaling Technology) were used. For flow cytometry, the following antibodies and dilutions were used: anti-CD31 (1:100) (#17-0319; eBioscience), anti-avb3 integrin (1:100) (MAB1976; Millipore), anti-VCAM-1(1:100) (#12-1069; eBioscience), anti-ICAM-1 (1:100) (#12-0549; eBioscience) and anti-MCP-1 (1:100) (#12-7099; eBioscience).

Faes S., Uldry E., Planche A., Santoro T., Pythoud C., Demartines N, & Dormond O. (2016). Acidic pH reduces VEGF-mediated endothelial cell responses by downregulation of VEGFR-2; relevance for anti-angiogenic therapies. Oncotarget, 7(52), 86026-86038.

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TNF-induced VCAM1 expression in MEFs

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MEFs were treated for 24 h with 10 ng/ml TNF, dissociated with trypsin/EDTA, and stained with anti-VCAM1 (eBioscience).

Mooster J.L., Le Bras S., Massaad M.J., Jabara H., Yoon J., Galand C., Heesters B.A., Burton O.T., Mattoo H., Manis J, & Geha R.S. (2015). Defective lymphoid organogenesis underlies the immune deficiency caused by a heterozygous S32I mutation in IκBα. The Journal of Experimental Medicine, 212(2), 185-202.

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Cytokine Regulation of Endothelial Markers

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The primary microvascular endothelial cells (HMVEC, Cat # CC-2527, Lonza Group, Ltd, USA) were obtained and were exposed to 0, 5 and 50 nM IL-18 for 24 hrs. These cells were stained with cell surface molecule-specific antibodies for flowcytometer analysis. The following reagents were used for specific antigen analysis: anti-ICAM-1, anti-VCAM-1 and anti-TNFα antibodies (eBiosciences). The cells were incubated for specific antigens with the required combination of antibodies at 4°C for 45 minutes followed by two washes. The flowcytometric analysis was performed using a FACSCalibur (BD Biosciences) and analyzed using Flow J software.

Niranjan R., Rajavelu P., Ventateshaiah S.U., Shukla J.S., Zaidi A., Mariswamy S.J., Mattner J., Fortgang I., Kowalczyk M., Balart L., Shukla A, & Mishra A. (2015). Involvement of Interleukin-18 in the pathogenesis of human eosinophilic esophagitis. Clinical immunology (Orlando, Fla.), 157(2), 103-113.

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Treg Cell Trafficking to Peripheral Tissues

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Mice were anaesthetized and 1 × 10⁶ Foxp3-GFP⁺CD25⁺ Tregs or activated Foxp3-GFP^-CD25⁺ no-Tregs were injected intradermally into the footpads or ear pinnae in 20 μL PBS as we previously described. For footpad assays, draining popliteal LNs were collected 12 hours post injection and processed for flow cytometry. For ear pinnae assays, whole-mount ears were collected 16 hours post injection. Sample were fixed for 10 minutes at room temperature with 4% paraformaldehyde, then permeabilized with 0.5% Triton X-100 for 30 min at 4°C; Samples were blocked for 1 hour in 5% donkey serum in PBS containing 0.1% Triton X-100 and incubated with listed antibodies overnight at 4 °C. Anti-Lyve-1 (1:100, Fitzgerald 70R-LR003), anti-CCL21 (1:100, R&D Systems), or anti-VCAM-1 (1:100, eBioscience 429). secondary antibodies were conjugated to Alexa Fluor 488, 546 or 647 (1:400, Jackson ImmunoResearch) for 1 hour at 4 °C, mounted in Prolong Gold with DAPI (Thermo Fisher, P36931). Distance of T cells from lymphatic vessels calculated with minimum distance program in Volocity 6.3.

Piao W., Xiong Y., Li L., Saxena V., Smith K.D., Hippen K.L., Paluskievicz C., Willsonshirkey M., Blazar B.R., Abdi R, & Bromberg J.S. (2020). Regulatory T Cells Condition Lymphatic Endothelia for Enhanced Transendothelial Migration. Cell reports, 30(4), 1052-1062.e5.

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Quantifying Cell Surface Expression

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To determine cell surface expression, 1x10 5 cells were cultured as indicated. Cells were stimulated with retinoic acid (100 nM), the agonistic anti-LTβR mAb or both for 24 hrs and 72 hrs. Cells were harvested by trypsinization and subsequently stained with biotin conjugated anti-VCAM-1 or anti-ICAM-1 (both eBioscience, Immunosource, Halle-Zoersel, Belgium), unlabeled anti-MAdCAM-1 (clone MECA 367, affinity purified from hybridoma cell culture supernatants) and unlabeled anti-LTβR mAb and subsequently counterstained with streptavidin-Alexa 488 and goat-anti-rat-Alexa 488 (all Invitrogen, Breda, The Netherlands), respectively. Sytox Blue (Invitrogen) staining was used to discriminate between live and dead cells. Data were acquired on a Cyan ADP High Performance Research Flow Cytometer (Beckman Coulter) and were analyzed with Summit Software v4.3.
For cell sorting, single cell suspensions of lymph nodes were stained in 100 µl diluted antibody mixtures containing the following antibodies; CD45-PE-Cy7 (Biolegend), Ter119-BV605 (Biolegend), GP38-Alexa Fluor 488 (Biolegend), CD31-PE (Biolegend), MHC-II-647(Clone M5-114, MO2Ab facility) and Cd11c Apc-Cy7 (Biolegend). Cells were sorted on a BD Fusion (BD Biosciences) using a 100 µm nozzle. Sorted cells were collected in ice-cold HBSS (Invitrogen), centrifuged and lysed in Trizol.

Koning J.J., Rajaraman A., Reijmers R.M., Konijn T., Pan J., Ware C.F., Butcher E.C., & Mebius R.E. (2020). Development of follicular dendritic cells in lymph nodes depends on retinoic acid mediated signaling.

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