PKI was purchased from Calbiochem and was used at 20μM for 1hr to inhibit PKA activity. The PECAM-1 antibody used for bead coating (PECAM 1.3, 5μg/25μl beads) was a generous gift from D.K. Newman (BloodCenter of Wisconsin). The RhoA antibody (26C4) used for western blotting (1:300) were purchased from Santa Cruz Biotechnologies. The phospho-CREB (Ser133), phospho-PKA (Tyr-197) and total PKA antibodies were from Cell Signaling and were used at 1:1000 dilution for western blotting, and the phospho-serine antibody (1:1000 dilution for western blot) and rhodamine-phalloidin (1:100 dilution for immunofluorescence) were from Invitrogen. The actin (1:2000 dilution for immunoblot), vinculin (1:100 for immunofluorescence) and β-catenin (1:100 for immunofluorescence) antibodies and GDP-β-S (20μM, 30min) were obtained from Sigma, and the HUTS-4 antibody (which recognizes ligated β1 integrin) was purchased from Millipore and used at 1:100 dilution for immunofluorescence. The VE-cadherin antibody was from B.D. Pharminogen and was used at 1:100 dilution.
Rhoa antibody 26c4
Manufactured by Santa Cruz Biotechnology
The RhoA antibody (26C4) is a tool used in research laboratories to detect and study the RhoA protein, a member of the Rho family of GTPases. This antibody is designed to specifically bind to and identify the RhoA protein, which plays a key role in regulating the organization and dynamics of the actin cytoskeleton. The antibody can be utilized in various techniques, such as Western blotting, immunoprecipitation, and immunocytochemistry, to investigate the expression, localization, and functions of RhoA in biological systems.
Automatically generated - may contain errors
Lab products found in correlation
Rhodamine phalloidin, by Thermo Fisher Scientific (2 mentions)
Gdp β s, by Merck Group (2 mentions)
Vinculin, by Merck Group (2 mentions)
β catenin, by Merck Group (2 mentions)
Phospho serine antibody, by Thermo Fisher Scientific (2 mentions)
Huts 4 antibody, by Merck Group (2 mentions)
Ve cadherin antibody, by BD (2 mentions)
Active rho pull down and detection kit, by Thermo Fisher Scientific (1 mentions)
3 protocols using rhoa antibody 26c4
1
Inhibition of PKA Regulates Endothelial Cell Signaling
2
Quantifying Active RhoA in VSMCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RhoA activation was assessed using an Active Rho Pull-Down and Detection Kit (Thermo Scientific) by immunoprecipitating GTP-bound RhoA with GST fusion protein of Rhotekin RhoA binding domain (GST-Rhotekin-RBD). Briefly, VSMCs plated in 150-mm dish were washed with ice-cold PBS and lysed in buffer containing 25 mM Tris pH 7.2, 150 mM NaCl, 5 mM MgCl2, 1% NP-40, and 5% glycerol. Cell lysates (500 μg) were incubated with 400 μg of GST-Rhotekin-RBD and glutathione resin at 4 °C for 1 h. Following washing, bound Rho was eluted by SDS sample buffer. The eluted samples and the total cell lysates were then subjected to Western blot analysis with RhoA antibody (26C4) (SantaCruz Biotechnology, sc-418) to detect active and total RhoA, respectively.
3
Inhibition of PKA Regulates Endothelial Cell Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
PKI was purchased from Calbiochem and was used at 20μM for 1hr to inhibit PKA activity. The PECAM-1 antibody used for bead coating (PECAM 1.3, 5μg/25μl beads) was a generous gift from D.K. Newman (BloodCenter of Wisconsin). The RhoA antibody (26C4) used for western blotting (1:300) were purchased from Santa Cruz Biotechnologies. The phospho-CREB (Ser133), phospho-PKA (Tyr-197) and total PKA antibodies were from Cell Signaling and were used at 1:1000 dilution for western blotting, and the phospho-serine antibody (1:1000 dilution for western blot) and rhodamine-phalloidin (1:100 dilution for immunofluorescence) were from Invitrogen. The actin (1:2000 dilution for immunoblot), vinculin (1:100 for immunofluorescence) and β-catenin (1:100 for immunofluorescence) antibodies and GDP-β-S (20μM, 30min) were obtained from Sigma, and the HUTS-4 antibody (which recognizes ligated β1 integrin) was purchased from Millipore and used at 1:100 dilution for immunofluorescence. The VE-cadherin antibody was from B.D. Pharminogen and was used at 1:100 dilution.
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!