Anti-LIMK1 Polyclonal Antibody is a Rabbit antibody targeting LIMK1. Anti-LIMK1 Polyclonal Antibody can be used in FCM,IF,IHC-Fr,IHC-P,WB.

IgG

Human,Mouse,Rat (predicted:Cow,Rabbit)

1. Tissue/cell: rat brain tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37°C for 20 min;
Incubation: Anti-LIMK1 Polyclonal Antibody, Unconjugated (TMAB-01070) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
2. Paraformaldehyde-fixed, paraffin embedded (mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (LIMK1) Polyclonal Antibody, Unconjugated (TMAB-01070) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
3. Paraformaldehyde-fixed, paraffin embedded (rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (LIMK1) Polyclonal Antibody, Unconjugated (TMAB-01070) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
4. Paraformaldehyde-fixed, paraffin embedded (Mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (LIMK1) Polyclonal Antibody, Unconjugated (TMAB-01070) at 1:400 overnight at 4°C, followed by a conjugated secondary antibody for 20 min and DAB staining.
5. Paraformaldehyde-fixed, paraffin embedded (Rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (LIMK1) Polyclonal Antibody, Unconjugated (TMAB-01070) at 1:400 overnight at 4°C, followed by a conjugated secondary antibody for 20 min and DAB staining.
6. Sample:
Spinal cord (Mouse) Lysate at 40 μg
Primary: Anti-LIMK1 (TMAB-01070) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 71 kDa
Observed band size: 75 kDa
7. Blank control (black line): Hela.
Primary Antibody (green line): Rabbit Anti-LIMK1 antibody (TMAB-01070)
Dilution: 1 μg/Test;
Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC
Dilution: 0.5 μg/Test.
Isotype control (orange line): Normal Rabbit IgG
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at-20°C, The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature.

WB: 1:500-2000; IHC-P: 1:20-100; IHC-Fr: 1:20-100; IF: 1:20-100; FCM: 1ug/Test

Polyclonal

KLH conjugated synthetic peptide: human LIMK1

Human

LIMK1

LIM domain kinase 2

Signal transduction,LIM,Neural Signal Transduction

Serine/threonine-protein kinase that plays an essential role in the regulation of actin filament dynamics. Acts downstream of several Rho family GTPase signal transduction pathways. Activated by upstream kinases including ROCK1, PAK1 and PAK4, which phosphorylate LIMK1 on a threonine residue located in its activation loop. LIMK1 subsequently phosphorylates and inactivates the actin binding/depolymerizing factors cofilin-1/CFL1, cofilin-2/CFL2 and destrin/DSTN, thereby preventing the cleavage of filamentous actin (F-actin), and stabilizing the actin cytoskeleton. In this way LIMK1 regulates several actin-dependent biological processes including cell motility, cell cycle progression, and differentiation. Phosphorylates TPPP on serine residues, thereby promoting microtubule disassembly. Stimulates axonal outgrowth and may be involved in brain development. Isoform 3 has a dominant negative effect on actin cytoskeletal changes.