Western blot analysis was performed as described previously.29 Briefly, aliquots of total protein (10 µg) were electrophoresed on sodium dodecyl sulfate polyacrylamide, 10% Tris‐HCl gels (Bio‐Rad Laboratories). The separated proteins were transferred to polyvinylidene difluoride membranes (Bio‐Rad Laboratories) and incubated with primary antibodies overnight at 4°C. Proteins were detected using the following antibodies: anti‐VE‐cadherin (Abcam), anti‐ZO‐1, anti‐E‐cadherin, anti‐Zinc Finger E‐Box Binding Homeobox 1 (ZEB1), anti‐N‐cadherin, anti‐Snail (Cell Signaling Technology, Danvers, MA, USA), anti‐CD13, anti‐CD133, anti‐EpCAM (Abcam) and anti‐β actin (Sigma, Tokyo, Japan).
Anti ve cadherin
Manufactured by Abcam
Sourced in United States, United Kingdom
Anti-VE-cadherin is a primary antibody product used in research applications. It targets the VE-cadherin (Vascular Endothelial-Cadherin) protein, which is a key component of endothelial cell-cell junctions. This antibody can be used to detect and analyze VE-cadherin expression and distribution in various experimental models.
Automatically generated - may contain errors
Lab products found in correlation
Anti cd31, by Abcam (9 mentions)
Anti occludin, by Abcam (8 mentions)
Anti zo 1, by Abcam (7 mentions)
Anti α sma, by Abcam (5 mentions)
Anti zo 1, by Thermo Fisher Scientific (5 mentions)
Anti α sma, by Merck Group (5 mentions)
Anti claudin 5, by Abcam (4 mentions)
Anti gapdh, by Abcam (4 mentions)
Anti p62, by Abcam (4 mentions)
Anti fsp 1, by Abcam (4 mentions)
Anti β actin, by Merck Group (3 mentions)
Anti caspase 3, by Abcam (3 mentions)
Anti beclin1, by Abcam (3 mentions)
Fluoview fv1000, by Olympus (3 mentions)
Anti zo 2, by Thermo Fisher Scientific (3 mentions)
Alexa fluor 488, by Thermo Fisher Scientific (3 mentions)
Anti fak, by Abcam (3 mentions)
Anti hif 1α, by Abcam (2 mentions)
Anti occludin, by Thermo Fisher Scientific (2 mentions)
Anti bax, by Abcam (2 mentions)
57 protocols using anti ve cadherin
1
Western Blot Protein Analysis Protocol
2
Investigating HIF-1α and Junctional Proteins in HCMECs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human cardiac microvascular endothelial cell (HCMECs) line (Cat 6110; ScienCell) was bought from ScienCell company and cultured in endothelial cell medium with 10% fetal bovine serum and maintained in 5% CO2 at 37°C in a humidified atmosphere. Cells were treated with TXL, CoCl2, pAd-GFP, or pAd-KLF4 for 24 hours and then harvested with lysis buffer containing 1% NP-40, 150 mM NaCl, 50 mM Tris-Cl pH 7.5, 10% glycerin, 1 mM Na3VO4, 1 mM phenylmethanesulfonyl fluoride (PMSF), and 1 mM dithiothreitol (DTT). The tissues were lysed with the above lysis buffer. Proteins were isolated from HCMECs, separated by sodium dodecyl sulfate–polyacrylamide gel electrophoresis, and transferred onto polyvinylidene difluoride membranes (Millipore). Membranes were blocked with 5% milk in Tris-HCl–Tween buffer solution for 2 hours at 37°C and incubated overnight at 4°C with specific antibodies [anti–HIF-1α (1:1000; Abcam), anti–VE-cadherin (1:1000; Abcam), anti–beta-catenin (1:10,000; Abgent), antioccludin (1:100,000; Epitomics), anti–claudin-1 (1:500; Novus), and anti-KLF4 (1:1000; Epitomics)]. After incubation with appropriate secondary antibodies, the membranes were developed using the chemiluminescence plus Western blot analysis kit (Millipore).
3
Proteomic analysis of colon tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Frozen colon tissue (~100 mg) was homogenized in cell lysis buffer (500 μl) (Cat EPX-99999-000 eBioscience, San Diego, USA) containing 0.1% proteinase inhibitor cocktail (Sigma, St. Louis, MO) and 1% NP40, in ceramic bead tubes using Fast Prep-24TM 5G machine (M.P. Biomedical LLC, California, USA) at a speed 6.0 ms for 30 s (twice). Samples were centrifuged at 12,000 g for 15 min at 4 °C, supernatants were collected and protein concentration was determined using Bio-Rad protein estimation kit (BioRad). 100 μg of protein was subjected to Western Blot analyses using 8% acrylamide gel. Membranes were probed with anti-pig primary antibodies raised in rabbit overnight at 4 °C and subsequently incubated with 1:10,000 dilution of goat-anti-rabbit HRP for 1 h at room temperature (BioRad Laboratories Inc., California). We used 1:1000 dilution of anti-VE-cadherin, anti-catenin, anti-HSP 27 (Abcam, Cambridge, MA) as primary antibodies. Detection was performed using a chemiluminescence system (super signal west chemiluminescent substrate, Thermo Scientific). Image Quant software (Image Quant TL 8.1 Version) was used for densitometric analysis. Anti-rabbit vinculin (Abcam ab73412) that cross reacts with pigs was used as a housekeeping protein for normalization of a target protein.
4
Investigating Signaling Pathways in Cellular Processes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The AKT inhibitor LY294002, GSK3β inhibitor SB216763, NFκB inhibitor Bay 11–7082, FITC-dextran, and Evans blue were purchased from Sigma (Sigma-Aldrich, St. Louis, MO, United States). JNK agonist anisomycin was purchased from Beyotime (Beyotime Biotechnology, Shanghai, China). TNF-α was obtained from R&D Systems (Minneapolis, MN, United States). The following primary antibodies were applied in this study: anti-p-GSK3β, anti-GSK3β, anti-p-JNK, anti-JNK, anti-p-NFκB and anti-NFκB antibodies purchased from Cell Signaling Technology (Danvers, MA, United States); anti-FGF20, anti-VE-cadherin, anti-Claudin-5, and anti-GFAP antibodies obtained from Abcam (Cambridge, MA, United States); and anti-p-AKT, anti-AKT and anti-Occludin antibodies purchased from Invitrogen (Carlsbad, CA, United States) and Santa Cruz Biotechnology (Dallas, TX, United States), respectively. The secondary antibodies used in this study were goat anti-rabbit immunoglobulin G (IgG) H&L (HRP) and goat anti-mouse IgG-HRP purchased from Abcam (Cambridge, MA, United States) and Santa Cruz Biotechnology (Dallas, TX, United States), respectively.
5
Immunofluorescence Analysis of Kidney and Endothelial Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Frozen kidney sections (5μm) and laminin-coated coverslips of HRGECs and HUVECs were used for immunofluorescence. Immunohistochemistry staining was performed as described (31 (link)). Primary antibodies used were as followed: Anti-LC3II (1:100; CST, USA), Anti-Beclin1 (1:100; CST, USA), Anti-CD31(1:100; Abcam, UK), Anti-α-SMA (1:100; Abcam, UK), Anti-VE-cadherin (1:100; Abcam, UK).
6
Comprehensive Lung Protein Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The total protein of lung tissues and cells was collected with RIPA buffer (Servicebio, Wuhan, China) with cocktail and phosphatase inhibitors (Thermo Fisher Scientific, Wuhan, China), followed by electrophoresed through 8–12% sodium dodecyl sulfate–polyacrylamide gel and then transferred to 0.45 μm PVDF membrane (Merck Millipore, USA). A total of 30 ug of protein was used for western blot experiments. After blocked with 5% milk, the membrane was immunoblotted with following primary antibodies(Except for labeled, all from Cell Signaling Technology, MA, USA) at 4 °C overnight: anti-ERRα (1:1000), anti-ZO-1 (1:1000; Abcam), anti-VE-cadherin (1:1000), anti-Occludin(1:1000), anti-JAM-A(1:1000; Abcam), anti-Sirt3 (1:1000), anti-Bcl-2 (1:1000), anti-Bax (1:1000), anti-Smac (1:1000), anti-Cytochrome c (1:1000), anti-caspase 3 (1:1000), anti-cleaved caspase 3 (1:500), anti-caspase 9 (1:1000), anti-cleaved caspase 9 (1:500), anti-LC3A/B(1:1000), anti-Beclin1 (1:1000), anti-p62 (1:1000), or anti-GAPDH (1:10000, Abcam, internal control). After incubated with the HRP-conjugated secondary antibodies at 25℃ for 1 h, the immunoblots were visualized using the ECL system. The gray value was analyzed using the Image J and calculate the relative protein level based on the density ratio of the target protein to GAPDH (internal control).
7
Western Blot Analysis of Endothelial Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Whole-cell lysates from 15,000 ECs were subjected to 8% gradient SDS-PAGE, and the proteins were transferred onto nitrocellulose membranes. Blots were blocked with 5% defatted milk and incubated with the appropriate primary antibodies overnight at 4 °C, followed by HRP-conjugated secondary antibodies at room temperature for 2 h. Membranes were developed using a chemiluminescence reagent (Thermo Fisher Scientific) [20 (link),31 (link)]. The primary antibodies were: anti-ICAM-1 (Cat# Ig60229-1-Ig, Proteintech, Chicago, IL, USA), anti-β-actin (Sigma-Aldrich Cat# A1978, RRID: AB_476,692), anti-VCAM-1 (Proteintech Cat# 11,444–1-AP, RRID: AB_2,214,232), anti-CD31 (BD Biosciences Cat# 560,983, RRID: AB_10,562,393), and anti-VE-cadherin (Abcam Cat# ab33168, RRID: AB_870,662).
8
Comprehensive Immune Cell Phenotyping in Pancreatitis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following antibodies were used for FACS staining, IHC staining and immunofluorescence staining: Anti-mouse CD4 BV650 (BioLegend 100546), anti-mouse-CD11b PerCP Cy55 (BioLegend, 101228), anti-mouse-Ly6G BV421 (BioLegend, 127628), anti-mouse-Ly6C BV605 (BioLegend 128036), anti-CD25-PECy7 (BioLegend, 102016), anti-CD69 BV510 (BioLegend, 104532), anti-CD8a BV605 (BioLegend, 100743), anti-FoxP3 APC (Miltenyi Biotec, 130-111-601), anti-mouse-CD11b (abcam, Ab133357), anti-mouse-CD68 (antibody-online, ABIN181836), anti-CCR2 (abcam, ab273050) anti-Ki67 (Bethyl, IHC-00375), anti-mouse-CD11b (abcam, Ab133357), anti-mouse-Ly6g (abcam, Ab25377), anti-mouse-Cystatin C (Novus biologicals, NB100-1033), anti-VE-cadherin (abcam, ab7047-50) anti-rabbit-HRP (DAKO, K4003), anti-mouse-HRP (DAKO, K4001). Caerulein was obtained from Sigma Aldricht (C9026-1MG, Munich, Germany), human myeloperoxidase from Calbiochem (Cat# 475911). All antibodies used in this study had been tested and established in previous studies (11 (link), 12 (link), 15 (link)). The concentration of the antibodies was used according to the manufacturer’s instructions.
9
Visfatin Induces Colocalization of Junctional Proteins in MVECs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For colocalization in MVECs, cells were grown on glass coverslips and then treated with 4 μg/ml visfatin (BioVision, Mountain View, CA, USA) for 24 hrs and was then terminated by the fixation of the cells in 4% (Paraformaldehyde) PFA for 15 min. as previously described 33 . Cells were washed in PBS and cells were incubated for 2 hrs at 4°C with rabbit anti‐ZO‐1 (1:500; Invitrogen, Grand Island, NY, USA), anti‐ZO‐2 (1:500; Invitrogen), anti‐Occludin (1:1000; Abcam, Cambridge, MA, USA), anti‐VE‐Cadherin (1:1000; Abcam). Double immunofluorescent staining was performed by Alexa Fluor 488 or Alexa Fluor 555‐labelled secondary antibody (1:100; Invitrogen) incubation for 1 hr at room temperature. The slices were visualized through sequentially scanning on an Olympus laser scanning confocal microscope (Fluoview FV1000; Olympus, Center Valley, PA, USA).
10
Immunohistochemical Analysis of SARS-CoV-2 Infection
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!