Rabbit LIMK1 Polyclonal Antibody | anti-LIMK1 antibody

Phospho-LIMK1 (Ser310) Antibody

Cat. Num. AAA9614280

• Gene Names: LIMK1; LIMK; LIMK-1
• Reactivity: Human, Mouse, Rat; Predicted Reactivity: Pig (100%), Bovine (100%), Horse (100%), Sheep (100%), Rabbit (100%), Dog (88%), Chicken (100%)
• Applications: Western Blot, Immunohistochemistry, Immunofluorescence, Immunocytochemistry, ELISA
• Purity: The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
• Synonyms: LIMK1; Polyclonal Antibody; Phospho-LIMK1 (Ser310) Antibody; EC 2.7.11.1; LIM domain kinase 1; LIM motif-containing protein kinase; LIMK; LIMK-1; limk1; LIMK1_HUMAN; anti-LIMK1 antibody

• Host: Rabbit
• Reactivity: Human, Mouse, Rat; Predicted Reactivity: Pig (100%), Bovine (100%), Horse (100%), Sheep (100%), Rabbit (100%), Dog (88%), Chicken (100%)
• Clonality: Polyclonal
• Isotype: Rabbit IgG
• Specificity: Phospho-LIMK1 (Ser310) Antibody detects endogenous levels of LIMK1 only when phosphorylated at Ser310.; Tissue Specificity: Highest expression in both adult and fetal nervous system. Detected ubiquitously throughout the different regions of adult brain, with highest levels in the cerebral cortex. Expressed to a lesser extent in heart and skeletal muscle.
• Purity/Purification: The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
• Form/Format: Liquid. Rabbit IgG in phosphate buffered saline, pH7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.
• Concentration: 1mg/ml (varies by lot)

• Applicable Applications for anti-LIMK1 antibody: Western Blot (WB), Immunohistochemistry (IHC), Immunofluorescence (IF), Immunocytochemistry (ICC), Peptide ELISA (EIA)
• Application Notes: WB: 1:1000-3000; IF/ICC: 1:100-1:500; IHC: 1:50-1:200; Peptide ELISA: 1:20,000-1:40,000
• Immunogen: A synthesized peptide derived from human LIMK1 around the phosphorylation site of Ser310.
• Conjugation: Unconjugated
• Fragment: Fab fragment
• Post Translational Modifications: Autophosphorylated. Phosphorylated on Thr-508 by ROCK1 and PAK1, resulting in activation. Phosphorylated by PAK4 which increases the ability of LIMK1 to phosphorylate cofilin. Phosphorylated at Ser-323 by MAPKAPK2 during activation of VEGFA-induced signaling, which results in activation of LIMK1 and promotion of actin reorganization, cell migration, and tubule formation of endothelial cells. Dephosphorylated and inactivated by SSH1. Phosphorylated by CDC42BP.Ubiquitinated. 'Lys-48'-linked polyubiquitination by RNF6 leads to proteasomal degradation through the 26S proteasome, modulating LIMK1 levels in the growth cone and its effect on axonal outgrowth. Also polyubiquitinated by RLIM (By similarity).
• Subunit Structure: Interacts (via LIM domain) with the cytoplasmic domain of NRG1. Interacts with NISCH. Interacts with RLIM and RNF6 (By similarity). Self-associates to form homodimers. Interacts with HSP90AA1; this interaction promotes LIMK1 dimerization and subsequent transphosphorylation. Interacts with CDKN1C. Interacts with SSH1. Interacts with ROCK1. Interacts (via LIM zinc-binding domains) with FAM89B/LRAP25 (via LRR repeat). Forms a tripartite complex with CDC42BPA, CDC42BPB and FAM89B/LRAP25 (By similarity).
• Similarity: Belongs to the protein kinase superfamily. TKL Ser/Thr protein kinase family.
• Subcellular Location: Cytoplasm. Nucleus. Cytoplasm>Cytoskeleton. Cell projection>Lamellipodium.; Note: Predominantly found in the cytoplasm. Localizes in the lamellipodium in a CDC42BPA, CDC42BPB and FAM89B/LRAP25-dependent manner.
• Preparation and Storage: Store at -20 degree C. Stable for 12 months from date of receipt.

Western Blot (WB)

(Western blot analysis of extracts from Raji cells, using Phospho-LIMK1 (Ser310) Antibody. The lane on the left was treated with blocking peptide.)

Western Blot (WB)

(Western blot analysis of LIMK1 (Phospho-Ser310) using PMA treated NIH-3T3 whole cell lysates.-/+ means absence or presence of N peptide（non-phospho peptide) and P peptide(phospho peptide).)

Immunohistochemistry (IHC)

(At 1/100 staining Rat brain tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)

Immunohistochemistry (IHC)

(At 1/100 staining Mouse brain tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)

Immunohistochemistry (IHC)

(At 1/100 staining Human ovarian cancer by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)

Immunohistochemistry (IHC)

(At 1/100 staining Human ovarian cancer and adjacent normal tissues by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)

Testing Data

(At 25 degree C. The primary antibody was diluted at 1/200 and incubated with the sample for 1 hour at 37 degree C. An Alexa Fluor 594 conjugated goat anti-rabbit IgG (H+L) Ab, diluted at 1/600, was used as the secondary antibody.)

Related Product Information for anti-LIMK1 antibody

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.Has a dominant negative effect on actin cytoskeletal changes. Required for atypical chemokine receptor ACKR2-induced phosphorylation of cofilin (CFL1).

NCBI and Uniprot Product Information

• NCBI GI #: 324715046
• NCBI GeneID: 3984
• NCBI Accession #: NP_001191355.1
• NCBI GenBank Nucleotide #: NM_001204426.1
• UniProt Accession #: P53667; O15283; Q15820; Q15821; Q75MU3; Q9Y5Q1; B7Z6I8; D3DXF4; D3DXF5
• Molecular Weight: 72,585 Da
• NCBI Official Full Name: LIM domain kinase 1 isoform 2
• NCBI Official Synonym Full Names: LIM domain kinase 1
• NCBI Official Symbol: LIMK1
• NCBI Official Synonym Symbols: LIMK; LIMK-1
• NCBI Protein Information: LIM domain kinase 1; LIM motif-containing protein kinase
• UniProt Protein Name: LIM domain kinase 1
• Protein Family: LIM domain kinase
• UniProt Gene Name: LIMK1
• UniProt Synonym Gene Names: LIMK; LIMK-1
• UniProt Entry Name: LIMK1_HUMAN

NCBI Description

There are approximately 40 known eukaryotic LIM proteins, so named for the LIM domains they contain. LIM domains are highly conserved cysteine-rich structures containing 2 zinc fingers. Although zinc fingers usually function by binding to DNA or RNA, the LIM motif probably mediates protein-protein interactions. LIM kinase-1 and LIM kinase-2 belong to a small subfamily with a unique combination of 2 N-terminal LIM motifs and a C-terminal protein kinase domain. LIMK1 is a serine/threonine kinase that regulates actin polymerization via phosphorylation and inactivation of the actin binding factor cofilin. This protein is ubiquitously expressed during development and plays a role in many cellular processes associated with cytoskeletal structure. This protein also stimulates axon growth and may play a role in brain development. LIMK1 hemizygosity is implicated in the impaired visuospatial constructive cognition of Williams syndrome. Alternative splicing results in multiple transcript variants encoding distinct isoforms.[provided by RefSeq, Feb 2011]

Uniprot Description

LIMK1: a TKL kinase of the LISK family which mediates Rho signaling to cytoskeleton. Contains two N-terminal LIM motifs. Phosphorylated and activated by PAK1 and ROCK, downstream effectors of Rho. May be involved in brain development. Phosphorylates and inactivates the actin binding/depolymerizing factor cofilin and induces actin cytoskeletal changes. The LIM domain interacts with the cytoplasmic domain of NRG1. Binds ROCK1. Interacts with slingshot 1. Overexpressed in prostate tumors and prostate and breast cancer cell lines; manipulation of activity correlates with invasiveness in breast and prostate cancer models. Located within the 7q11.2 amplicon associated with metastatic prostate cancer. Loss of one copy of LIMK1 is the likely cause of visuospatial cognition defects seen in small chromosomal deletions associated with dominant Williams-Beuren syndrome Three splice variant have been identified.

Protein type: Protein kinase, TKL; Kinase, protein; EC 2.7.11.1; Protein kinase, Ser/Thr (non-receptor); TKL group; LISK family; LIMK subfamily

Chromosomal Location of Human Ortholog: 7q11.23

Cellular Component: nucleoplasm; neuron projection; focal adhesion; cytoplasm; cytosol

Molecular Function: protein serine/threonine kinase activity; protein binding; zinc ion binding; protein heterodimerization activity; heat shock protein binding; ATP binding; protein kinase activity

Biological Process: axon guidance; nervous system development; positive regulation of axon extension; positive regulation of stress fiber formation; negative regulation of ubiquitin-protein ligase activity; ephrin receptor signaling pathway; innate immune response; positive regulation of actin filament bundle formation; actin cytoskeleton organization and biogenesis; signal transduction; protein amino acid phosphorylation; Rho protein signal transduction

Disease: Williams-beuren Syndrome