All lysis buffers were supplemented with 1% protease inhibitor (Sigma Aldrich, USA). Protein extracts were resolved using gradient precast SDS – polyacrylamide gel electrophoresis (Biorad Laboratories Inc, USA), and then electro-transferred onto a nitrocellulose membrane by using Trans-Blot® Turbo™ Transfer System (Biorad Laboratories Inc, USA) for immunoblot analysis. Antibody probing was done as per manufacturers’ instructions Antibodies used include anti-RhoA antibody (1:1000, Abcam, UK), anti-CDC42 antibody (1:1000, Abcam, UK), anti-Rac1 antibody (1:1000 Abcam, UK), anti-phospho-MYPT1 (Thr696) antibody (1:1000, Cell Signaling Technology, USA), anti-myosin light chain (phospho S20) antibody (1:1000, Abcam, UK), anti-GAPDH antibody (1:2000, Abcam, UK). Secondary antibodies either IRDye® 800CW Goat anti-Mouse IgG (LI-COR, USA) or IRDye® 800CW Goat anti-Rabbit IgG (LI-COR, USA) were used with dilution of 1:15000. Specific protein bands were detected using Odyssey® CLx Imaging System (LI-COR, USA).
Anti cdc42 antibody
Manufactured by Abcam
Sourced in United Kingdom
Anti-CDC42 antibody is a laboratory reagent used for the detection and quantification of the CDC42 protein in various biological samples. CDC42 is a small Rho GTPase that plays a crucial role in regulating cellular processes such as cell division, migration, and cytoskeletal organization. This antibody can be used in techniques like Western blotting, immunohistochemistry, and immunoprecipitation to study the expression and localization of CDC42 in cells and tissues.
Automatically generated - may contain errors
Lab products found in correlation
Odyssey clx imaging system, by LI COR (1 mentions)
Anti rhoa antibody, by Abcam (1 mentions)
Gradient precast sds polyacrylamide gel electrophoresis, by Bio-Rad (1 mentions)
Anti phospho mypt1 thr696 antibody, by Cell Signaling Technology (1 mentions)
Anti rac1 antibody, by Abcam (1 mentions)
Irdye 800cw goat anti mouse igg, by LI COR (1 mentions)
Irdye 800cw goat anti rabbit igg, by LI COR (1 mentions)
Protease inhibitor, by Merck Group (1 mentions)
Trans blot turbo transfer system, by Bio-Rad (1 mentions)
Anti gapdh antibody, by Abcam (1 mentions)
Phalloidin ifluor 488, by Abcam (1 mentions)
Oil red o solution, by Solarbio (1 mentions)
Biotin labeled sna, by Vector Laboratories (1 mentions)
Er tracker blue white dpx, by Yeasen (1 mentions)
Biotin labeled aal, by Vector Laboratories (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Low glucose dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions)
Dmem containing 4.5 g l glucose, by Thermo Fisher Scientific (1 mentions)
Elisa kit for insulin, by Cloud-Clone (1 mentions)
5 protocols using anti cdc42 antibody
1
Immunoblot Analysis of Rho GTPases
2
Endothelial Dysfunction and CDC42 Regulation
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Ox-LDL was purchased from Yiyuanbiotech (YB-002, Guangzhou, China). The anti-CDC42 antibody, anti-Rac1/2/3 antibody, anti-PAK antibody, anti-pCDC42 antibody, anti-pPKA antibody, and Phalloidin-iFluor 488 were purchased from Abcam (Oxford, UK). The anti-Unc5b antibody was purchased from CST (Danvers, USA). The PAK inhibitor (FRAX486) and CDC42 inhibitor (ML141) were purchased from MCE (MCE, USA). Biotin-labeled LTL, biotin-labeled LCA, biotin-labeled AAL, biotin-labeled UEA1, biotin-labeled SNA, biotin-labeled MAL1, biotin-labeled VVL, and biotin-labeled PHA-L were purchased from Vector Laboratories (Peterborough, UK). ER-Tracker Blue‒White DPX was purchased from Yeasen Biotech Co., Ltd. (Shanghai, China). Oil Red O solution was purchased from Solarbio Science & Technology Co., Ltd. (Beijing, China). FBS was purchased from BI (Israel).
3
Wnt/Cdc42 Signaling in Rat ADSC Differentiation
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Anti-Cdc42 antibody was purchased from Abcam (Cambridge, MA, USA) and anti-Dvl-2 antibody was purchased from Cell Signaling Technology (Trask Lane Danvers, MA, USA). Anti-Insulin, anti-Ngn3, anti-NeuroD1, anti-PDX1, anti-non-p-GSK3β, anti-p-GSK3β, anti-non-p-β-catenin, and anti-β-actin antibodies were purchased from Affinity Biosciences (Cincinnati, OH, USA). HPR-labeled anti-rabbit IgG of goat and HPR-labeled anti-rat IgG of goat were purchased from Zhongshan Jinqiao Company (Beijing, China). Recombinant human Wnt-3a was purchased from R&D Inc. (Minneapolis, MN, USA), and Cdc42 inhibitor ML141 (5mg) was purchased from Selleck (Houston, TX, USA). DTZ was obtained from Sigma (St. Louis, MO, USA). Bovine serum albumin (BSA), Triton-100 and DMSO were purchased from Solarbio (Beijing, China). Low-glucose Dulbecco’s modified Eagle’s medium (DMEM) and DMEM containing 4.5 g/L glucose were obtained from Gibco-BRL (Gaithersburg, MD, USA). Wistar rat ADSC basal medium was purchased from Cyagen (USA). Fetal bovine serum (FBS) was purchased from Biological Industries (Cromwell, CT, USA). Penicillin and streptomycin were purchased from Gibco Life Technologies (Carlsbad, CA, USA). Krebs-Ringer bicarbonate HEPES (KRBH) was purchased from PanEra Company (Guangzhou, China). Enzyme-linked immunosorbent assay (ELISA) Kit for Insulin was purchased from Cloud-Clone Corp (Wuhan, China).
4
Enrichment of Active Cdc42 from Cells
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Cells were cultured for 96 h in 3D collagen matrices, and GTP-loaded Cdc42 from cell extracts was enriched by pull down with immobilized PBD domains from Pak1 (Kutys and Yamada, 2014 (link)). Briefly, endothelial cells were lysed by sonication in ice-cold lysis buffer containing 20 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), pH 7.6, 1% Triton X-100, 150 mM NaCl, 5 mM MgCl2, 1 mM dithiothreitol (DTT), Sigma-Aldrich protease and phosphatase inhibitor cocktail, and 5% glycerol. After high-speed centrifugation for 10 min at 4°C, supernatants were collected and protein concentration determined by the Bradford method. Next 500 μg of protein was incubated in the presence of 40 μg of PBDPak1–glutathione S-transferase protein beads (Cytoskeleton) for 1 h at 4°C. Beads were washed twice with 500 μl of ice-cold wash buffer (25 mM Tris-HCl, pH 7.5, 30 mM MgCl2, 40 mM NaCl, 1 mM DTT, 1 mM phenylmethylsulfonyl fluoride (PMSF), 5 μg/ml aprotinin, and 1 μg/ml leupeptin) and boiled in sample buffer (40 μl of beads in wash buffer plus 10 μl of 5× sample buffer) for 5 min. Samples were resolved by SDS–PAGE, followed by Western blotting. Membranes were probed with an anti-Cdc42 antibody (Abcam).
5
Molecular Profiling of Oocyte Maturation
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About 300 oocytes of each group were briefly washed in phosphate-buffered saline and then lysed in 2× SDS buffer and stored at −80°C until use. Immunoblot analysis was preformed as described (Huang et al., 2015 (link)). For immunolabeling, the following primary antibodies and dilutions were used: goat anti–SPAG-1 antibody (1:200), anti–phospho-AMPKα antibody (1:1500), anti–phospho-cyclin B1 antibody (1:1000), anti–histone H3 (acetyl K9) antibody (1:1000), rabbit anti–cyclin B2 polyclonal antibody (1:200; Santa Cruz Biotechnology), rabbit anti–α-tubulin monoclonal antibody (1:1000; Cell Signaling Technology), rabbit anti–acetylated-α-tubulin (1:1000; Cell Signaling Technology), anti–phospho-H2A.X antibody (1:1000; Cell Signaling Technology), rabbit anti–phospho-p44/42 MAPK antibody (1:1000), rabbit anti-PKC monoclonal antibody (1:1000; Abcam), anti-CDC42 antibody (1:1000; Abcam), anti–N-WSAP antibody (1:1000; Abcam), anti-Arp3 antibody (1:1000; Abcam), and anti-BubR1 antibody (1:1000). The data were normalized to β-actin.
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