Product datasheet
Anti-LIMK1 antibody ab81046
2 References 3 Images
Overview
Product name
Anti-LIMK1 antibody
Description
Rabbit polyclonal to LIMK1
Specificity
This antibody is specific for Limk1 and does not detect Limk2
Tested applications
WB, ICC/IF, IHC-P
Species reactivity
Reacts with: Rat, Human
Predicted to work with: Mouse, Orangutan
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 600 to the C-terminus of
Human LIMK1.Read Abcam's proprietary immunogen policy(Peptide available as ab90604.)
Positive control
This antibody gave a positive result in the following lysates: Human brain tissue; Rat cortex
tissue lysate; SHSY-5Y; SK N BE.
Properties
Form
Liquid
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or 80°C. Avoid freeze / thaw cycle.
Storage buffer
Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
Applications
Our Abpromise guarantee covers the use of ab81046 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application
WB
Abreviews
Notes
Use a concentration of 1 µg/ml. Detects a band of approximately 73 kDa
(predicted molecular weight: 73 kDa).
ICC/IF
Use a concentration of 1 µg/ml.
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Application
IHC-P
Abreviews
Notes
Use a concentration of 5 µg/ml.
Target
Function
Protein kinase which regulates actin filament dynamics. Phosphorylates and inactivates the actin
binding/depolymerizing factor cofilin, thereby stabilizing the actin cytoskeleton. Stimulates axonal
outgrowth and may be involved in brain development. Isoform 3 has a dominant negative effect
on actin cytoskeletal changes.
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.
Involvement in disease
Note=LIMK1 is located in the Williams-Beuren syndrome (WBS) critical region. WBS results
from a hemizygous deletion of several genes on chromosome 7q11.23, thought to arise as a
consequence of unequal crossing over between highly homologous low-copy repeat sequences
flanking the deleted region.
Sequence similarities
Belongs to the protein kinase superfamily. TKL Ser/Thr protein kinase family.
Contains 2 LIM zinc-binding domains.
Contains 1 PDZ (DHR) domain.
Contains 1 protein kinase domain.
Post-translational
modifications
Autophosphorylated.
Phosphorylated on serine and/or threonine residues by ROCK1. May be dephosphorylated and
inactivated by SSH1.
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.
Cellular localization
Cytoplasm. Cell projection > growth cone.
Anti-LIMK1 antibody images
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All lanes : Anti-LIMK1 antibody (ab81046) at
1 µg/ml
Lane 1 : Brain (Human) Tissue Lysate - adult
normal tissue (ab29466)
Lane 2 : Rat Cortex Tissue Lysate
Lane 3 : SHSY-5Y (Human neuroblastoma
cell line) Whole Cell Lysate
Lane 4 : SK N BE (Human neuroblastoma)
Whole Cell Lysate
Western blot - LIMK1 antibody (ab81046)
Lysates/proteins at 10 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG - H&L - PreAdsorbed (HRP) at 1/3000 dilution
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 73 kDa
Observed band size : 73 + 75 kDa
Additional bands at : 22 kDa,87 kDa. We
are unsure as to the identity of these extra
bands.
Exposure time : 20 minutes
ICC/IF image of ab81046 stained PC12 cells.
The cells were 4% PFA fixed (10 min) and
then incubated in 1%BSA / 10% normal Goat
serum / 0.3M glycine in 0.1% PBS-Tween for
1h to permeabilise the cells and block nonspecific protein-protein interactions. The cells
were then incubated with the antibody
(ab81046, 1µg/ml) overnight at +4°C. The
secondary antibody (green) was Alexa Fluor®
Immunocytochemistry/ Immunofluorescence -
488 Goat anti-Rabbit IgG (H+L) used at a
LIMK1 antibody (ab81046)
1/1000 dilution for 1h. Alexa Fluor® 594 WGA
was used to label plasma membranes (red) at
a 1/200 dilution for 1h. DAPI was used to
stain the cell nuclei (blue) at a concentration of
1.43µM.
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IHC image of ab81046 staining in Human
Hippocampus formalin fixed paraffin
embedded tissue section, performed on a
Leica BondTM system using the standard
protocol F. The section was pre-treated using
heat mediated antigen retrieval with sodium
citrate buffer (pH6, epitope retrieval solution
1) for 20 mins. The section was then
incubated with ab81046, 5µg/ml, for 15 mins
Immunohistochemistry (Formalin/PFA-fixed
at room temperature and detected using an
paraffin-embedded sections) - LIMK1 antibody
HRP conjugated compact polymer system.
(ab81046)
DAB was used as the chromogen. The
section was then counterstained with
haematoxylin and mounted with DPX.
For other IHC staining systems (automated
and non-automated) customers should
optimize variable parameters such as antigen
retrieval conditions, primary antibody
concentration and antibody incubation times.
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