Cells were fixed with 4% paraformaldehyde (PFA) in PBS for 10 minutes before permeabilisation in PBS containing 0.2% Triton-X for 10 minutes at room temperature. The cells were stained with DAPI (Sigma, d9542, MO, USA) to reveal DNA in cell nuclei and TRITC-phalloidin (Sigma, p1951, MO, USA) to reveal F-actin in 3% bovine serum albumin with PBS + 0.1% Tween-20. Cells were stained using anti-pMLC (S19) (Cell signaling technology #3675) or anti-pERM antibody (Cell signaling technology #3141) followed by appropriate Alexafluor-conjugated secondary antibodies (Invitrogen). PDPN-V5 was stained using anti-V5 FITC-conjugated antibody (Invitrogen R963-25). ARP2/3 was stained using Anti-p34-Arc/ARPC2 antibody (07–227 Millipore).
Anti perm antibody
Manufactured by Cell Signaling Technology
Sourced in United States
The Anti-pERM antibody is a lab equipment product that detects phosphorylated Ezrin/Radixin/Moesin (pERM) proteins. It is a useful tool for researchers studying the activation and regulation of ERM proteins, which play important roles in cell cytoskeleton organization and signal transduction.
Automatically generated - may contain errors
Lab products found in correlation
Anti v5 fitc conjugated antibody, by Thermo Fisher Scientific (2 mentions)
Anti p34 arc arpc2 antibody, by Merck Group (2 mentions)
Anti pmlc s19, by Cell Signaling Technology (2 mentions)
Phalloidin tritc, by Merck Group (2 mentions)
Alexa fluor conjugated secondary antibody, by Thermo Fisher Scientific (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (2 mentions)
Anti h2a x antibody, by Cell Signaling Technology (1 mentions)
Anti alpha tubulin antibody, by Thermo Fisher Scientific (1 mentions)
Anti lc3 antibody, by Cell Signaling Technology (1 mentions)
Anti cdx2 antibody, by BioGenex (1 mentions)
Anti nanog antibody, by Abcam (1 mentions)
Anti p erk1 2 antibody, by Cell Signaling Technology (1 mentions)
Anti moesin antibody, by Cell Signaling Technology (1 mentions)
Nitrocellulose membrane, by Merck Group (1 mentions)
Anti erk1 2 antibody, by Cell Signaling Technology (1 mentions)
Anti β actin antibody, by Merck Group (1 mentions)
Anti α sma antibody, by Merck Group (1 mentions)
4 protocols using anti perm antibody
1
Cytoskeletal Protein Localization Assay
2
Antibody Sourcing and Characterization
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All chemicals and drugs were purchased from Sigma (St. Louis, MO, USA) unless otherwise indicated. The anti-H2A.X antibody, anti-LC3 antibody, anti-rabbit IgG (H + L), F (ab’)2 Fragment (Alexa Fluor® 594 Conjugate) antibody, anti-pERM antibody, anti-pMRLC antibody were purchased from Cell Signaling Technology (Danvers, MA, USA). The anti-CDX2 antibody was purchased from BioGenex (San Francisco, USA). The anti-Nanog antibody was purchased from Abcam (Cambridge, United Kingdom). The anti-alpha Tubulin antibody was purchased from Thermo Fisher (Shanghai, China). The Fluorescein (FITC)–conjugated Affinipure Goat Anti-Rabbit IgG (H + L) secondary antibody was purchased from Proteintech (Beijing, China).
3
Cytoskeletal Protein Localization Assay
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Cells were fixed with 4% paraformaldehyde (PFA) in PBS for 10 minutes before permeabilisation in PBS containing 0.2% Triton-X for 10 minutes at room temperature. The cells were stained with DAPI (Sigma, d9542, MO, USA) to reveal DNA in cell nuclei and TRITC-phalloidin (Sigma, p1951, MO, USA) to reveal F-actin in 3% bovine serum albumin with PBS + 0.1% Tween-20. Cells were stained using anti-pMLC (S19) (Cell signaling technology #3675) or anti-pERM antibody (Cell signaling technology #3141) followed by appropriate Alexafluor-conjugated secondary antibodies (Invitrogen). PDPN-V5 was stained using anti-V5 FITC-conjugated antibody (Invitrogen R963-25). ARP2/3 was stained using Anti-p34-Arc/ARPC2 antibody (07–227 Millipore).
4
Western Blot Analysis of Epithelial-Mesenchymal Markers
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HK-2 cells were lysed in sample buffer containing RIPA and phenylmethanesulfonyl fluoride (PMSF). Protein concentrations were determined by BCA protein assay kit (Pierce). Thirty micrograms of protein extract was separated by SDS-PAGE gel and transferred onto nitrocellulose membranes (Millipore). The blots were blocked with 5% nonfat dry milk in Tris-buffered saline containing 0.05% Tween 20 (TTBS) at room temperature for 1 hr, followed by incubation with antibodies against primary antibodies at 4°C overnight. After three washes, the membrane was then incubated with horseradish peroxidase (HRP)-conjugated secondary antibodies (Kirkegaard & Perry Laboratories) and visualized with ECL kits (GE Amersham). The primary antibodies that were used included anti-α-SMA antibody(1∶1,000, Sigma), anti-E-cadherin antibody (1∶500, BD Pharmigen), anti-moesin antibody (1∶1000, Cell Signaling), anti-Erk 1/2 antibody (1∶1000, Cell Signaling), anti-pErk 1/2 antibody (1∶1000, Cell Signaling), anti-pERM antibody (1∶1000, Cell Signaling) and anti-β-actin antibody (1∶10000, Sigma Aldrich).
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