Esculin (≥98%, HPLC) was purchased from Aladdin Chemical Reagent Co., Ltd. (Shanghai, China). Protein G-Sepharose™ 4B beads, LPS, fMLP and calcein-AM was obtained from Sigma-Aldrich (St. Louis, MO). Recombinant human intercellular adhesion molecules 1 (rhICAM-1) or human IgG were purchased from R&D Systems (Abingdon, VA). ELISA kits for TNF-α, IL-6 and IL-1β were obtained from Invitrogen (Vienna, Austria). ELISA kit for MCP-1 was purchased from Beyotime Biotechnology (Shanghai, China). Antibodies against IκBα (ab76429), p-IκBα (ab133462), Vav1 (ab97574) and phospho-Vav1 (ab76225) were acquired from Abcam (Cambridge, MA). Anti-PAK1 (#2602), anti-phospho-PAK1 (#2601), anti-LIMK1 (#3842), anti-phospho-LIMK1(#3841), anti-cofilin (#3318), anti-phospho-cofilin (#3311), anti-p65 (#8242), anti-p-p65 (#3033), anti-GST (#2624), anti-Rac1 (#4651), anti-ICAM-1 (#4915), anti-β2 integrin (#73663) and anti-β-actin (#4970) antibodies were purchased from Cell Signaling Technology (Berkeley, CA). Enhanced Supersignal West Pico Chemiluminescence (ECL) substrate kit was purchased from Thermo Scientific (Rockford, IL).
Ab97574
Manufactured by Abcam
Sourced in China
Ab97574 is a laboratory equipment product. It is a multipurpose instrument designed for various laboratory applications. The core function of this product is to perform tasks required in a research or analytical setting. Further details about the specific capabilities and intended use of this product are not available.
Automatically generated - may contain errors
Lab products found in correlation
Lipopolysaccharides (lps), by Merck Group (1 mentions)
Calcein am, by Merck Group (1 mentions)
Formyl methionyl leucyl phenylalanine (fmlp), by Merck Group (1 mentions)
Ab76429, by Abcam (1 mentions)
Anti β actin, by Cell Signaling Technology (1 mentions)
Rhicam 1, by R&D Systems (1 mentions)
Protein g sepharose 4b beads, by Merck Group (1 mentions)
Ab76225, by Abcam (1 mentions)
Ab133462, by Abcam (1 mentions)
Imagequant las 4000 mini, by Fujifilm (1 mentions)
Pierce bca protein assay kit, by Thermo Fisher Scientific (1 mentions)
Eblot l1 wet protein transfer system, by GenScript (1 mentions)
Af7018, by Affinity Biosciences (1 mentions)
Ab273023, by Abcam (1 mentions)
3 protocols using ab97574
1
Esculin Inhibits Inflammatory Responses
2
Western Blot Analysis of Immune Proteins
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Lung tissues were minced and lysed in ice-cold RIPA Lysis Buffer containing phosphatase inhibitors and a protease inhibitor to obtain protein. Protein concentrations were quantified with Pierce BCA Protein Assay Kit (Thermo Scientific). For western blot, 30 μg of protein was loaded in each well and separated by 10% SDS-PAGE, and then, the protein bands were electro-transferred onto 0.2 μm PVDF membranes by the eBlot™ L1 wet protein transfer system (GenScript). The blocked blots were incubated with anti-CD11α (anti-ITGAL, 1 : 1000, ab228964, Abcam), anti-Syk (1 : 1000, 13198T, Cell Signaling Technology), anti-Vav1 (1 : 1000, ab97574, Abcam), and anti-beta actin (1 : 3000, AF7018, Affinity) at 4°C overnight, followed by incubation with HRP-conjugated secondary antibodies (1 : 10000) for 1 h. The blots were visualized using ImageQuant LAS-4000 mini (Fujifilm Corporation, Tokyo, JP) and then quantified by the ImageJ software.
3
Immunohistochemical Protein Expression Analysis
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The paraffin-embedded tissue blocks were sectioned at 3-μm thickness. IHC was performed using a DAB kit (GK600710, Shanghai, China) following the manufacturer’s protocol. The anti-VAV1 antibody (ab97574, 1:100), anti-IL2RG antibody (ab273023, 1:50), anti-CD93 antibody (ab198854, 1:100), and anti-CD62P antibody (ab182135, 1:300) used for IHC was purchased from Abcam (Shanghai, China). Goat anti-rabbit (ZF-0513, Zhongshan Golden Bridge, Beijing, China) as a secondary antibody. The NanoZoomer S210 Digital slide scanner scanned the image. Five randomly selected fields per section per sample at 400x magnification. Image-Pro Plus 6.0 was used to determine integrated optical density (IOD) values, and the IOD per unit area (mean density) represented the specific protein relative expression level. Statistical analyses were performed using GraphPad Prism (version 8.0.1).
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