Cultured cells were exposed to isoflavones or E2 for 30 min then rinsed three times with PBS, fixed with 4% PFA, and blocked with 2% FBS. Cells were then incubated with rabbit monoclonal anti-phospho-Akt (Ser473) (D9E) XP (1:200; Cell Signaling, MA, USA), anti-phospho-p44/42 MAPK (ERK1/2) (Thr202/Tyr204) (1:200; Cell Signaling), or anti-phospho-Rac1/Cdc42 (Ser71) (1:200; Cell Signaling) antibodies, followed by CytoPainter Phalloidin-iFluor 594 reagent (Abcam) and donkey anti-rabbit IgG (H+L) secondary antibodies, Alexa Fluor® 488 conjugate (1:200; Thermo Fisher Scientific, Inc, Waltham, MA, USA). Cell nuclei were also stained with DAPI. The cells were then inspected using a laser confocal scanning microscope (Zeiss LSM 880, Carl Zeiss Microscopy GmbH).
Anti phospho rac1 cdc42 ser71
Manufactured by Cell Signaling Technology
Sourced in United States
The Anti-phospho-Rac1/Cdc42 (Ser71) is a primary antibody that recognizes the phosphorylated forms of the Rac1 and Cdc42 proteins at serine 71. This antibody can be used to detect the activated states of these small GTPases.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 880, by Zeiss (1 mentions)
Alexa fluor 488 conjugate, by Thermo Fisher Scientific (1 mentions)
Cytopainter phalloidin ifluor 594 reagent, by Abcam (1 mentions)
Anti phospho akt ser473 d9e xp, by Cell Signaling Technology (1 mentions)
Anti phospho p44 42 mapk erk1 2 thr202 tyr204, by Cell Signaling Technology (1 mentions)
Pvdf membrane, by Cytiva (1 mentions)
Anti pak1 2 3, by Cell Signaling Technology (1 mentions)
Anti gapdh, by Cell Signaling Technology (1 mentions)
Hrp conjugated anti rabbit secondary antibody, by Cell Signaling Technology (1 mentions)
Anti arp2, by Cell Signaling Technology (1 mentions)
Anti rac1 cdc42, by Cell Signaling Technology (1 mentions)
2 protocols using anti phospho rac1 cdc42 ser71
1
Isoflavone and E2 Signaling Pathways
2
Western Blot Analysis of Actin Regulators
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Whole cell protein extracts were obtained by direct lysis in Lammelli's Buffer and boiled at 100°C for 10 min. Proteins were separated on 12% SDS-polyacrylamide gels and transferred to PVDF membranes (Amersham Biosciences, UK). Non-specific binding sites were blocked for 1 hour at room temperature with 5% non-fat dry milk in TBS-T. Membranes were incubated overnight at 4°C with anti-Rac1/Cdc42 (#4651), anti-Phospho-Rac1/Cdc42 (Ser71) (#2461), anti-ARP2 (#3128), anti-PAK1/2/3 (#2604), anti-Phospho-PAK1 (Thr423)/PAK2 (Thr402) (#2601), and anti-GAPDH (#2118) (Cell Signalling, Leiden, The Netherlands). All primary antibodies were used at 1:1000, washed 3 times with TBS-T, followed by incubation with HRP-conjugated anti-rabbit secondary antibodies (#7074, 1:1000, Cell Signalling, Leiden, The Netherlands). Protein complexes were visualized with a chemiluminescent detection kit (Novex ECL) and bands were quantified using ImageJ software.
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!