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Mouse anti vcam 1 monoclonal antibody

Manufactured by Abcam
Sourced in United Kingdom, United States

The Mouse anti-VCAM-1 monoclonal antibody is a laboratory reagent used in research applications. It recognizes and binds to the Vascular Cell Adhesion Molecule 1 (VCAM-1), which is a cell surface protein involved in cell-cell adhesion. This antibody can be utilized in various immunological techniques to study the expression and function of VCAM-1 in biological systems.

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2 protocols using mouse anti vcam 1 monoclonal antibody

1

Evaluating VCAM-1 Expression on HUVECs

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Expression of VCAM-1 on the surface of HUVECs was evaluated by incubating 2 × 105 HUVECs in EGM in the absence or presence of 20 ng ml−1 human TNFα (hTNFα Millipore, Billerica, MA, USA) for 24 h. After harvesting and washing with phosphate-buffered saline solution (PBS), the cells were fixed in 4% paraformaldehyde (PFA) for 20 min at room temperature. After blocking with PBS containing 1% bovine serum albumin (BSA) for 1 h at room temperature, the cells were incubated first with mouse anti-VCAM-1 monoclonal antibody (1 μg ml−1; Abcam, Cambridge, UK) for 1 h at 37 °C, and then with Alexa Fluor 488-conjugated anti-mouse antibody (1:1000; Invitrogen) for 1 h at 37 °C. The effects of anti-VCAM-1-D6 IgG on endothelial cell activation were evaluated by incubating 2 × 105 HUVECs in the absence or presence of 20 ng ml−1 hTNFα (Millipore), 20 μg ml−1 control IgG or anti-VCAM-1-D6 IgG for 24 h. After blocking with PBS containing 1% BSA for 1 h at room temperature, the cells were incubated first with rabbit anti-ICAM-1 monoclonal antibody (1:500; Abcam) for 1 h at 37 °C, and then with an Alexa Fluor 488-conjugated anti-rabbit antibody (1:1000; Invitrogen) for 1 h at 37 °C. Samples were analyzed by flow cytometry using a FACSCalibur system (BD Biosciences, San Jose, CA, USA) with the aid of FlowJo software (TreeStar, Ashland, OR, USA).
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2

Quantitative Immunohistochemistry of VCAM-1 in Lung

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Immunohistochemistry was performed as described previously with minor modifications [23 (link)]. Briefly, tissue slides printed with normal lung or lung cancer tissues were purchased from SuperBioChips Laboratories (Seoul, Korea). The slides were incubated first with mouse anti-VCAM-1 monoclonal antibody (1:200; Abcam, Cambridge, MA, USA) and then with biotinylated goat anti-mouse IgG (1:200; Vector Laboratories, Burlingame, CA, USA). Immunoreactive proteins were visualized using VECTASTAIN ABC Reagent (Vector Laboratories). For chromogenic reactions, the slides were incubated with fresh 3,3′-diaminobenzidine tetrahydrochloride solution (Vector Laboratories). All samples were counterstained with Meyer’s hematoxylin (Vector Laboratories). VCAM-1 expression was observed by light microscopy using an Olympus BX51 microscope (Olympus, Tokyo, Japan), and RGB images were acquired using Paint Shop Pro X software (Corel, Ottawa, ON, Canada). After performing background subtraction, VCAM-1 expression was quantified by measuring the signal density with Image J software version 1.48v (National Institutes of Health, Bethesda, MA, USA).
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