Overview

  • Product name

    Anti-LIM Kinase 1 (phospho T508) antibody
    See all LIM Kinase 1 primary antibodies
  • Description

    Rabbit polyclonal to LIM Kinase 1 (phospho T508)
  • Host species

    Rabbit
  • Specificity

    This antibody is specific for LIM Kinase only when phosphorylated at threonine 508. It recognizes both LIMK1 and LIMK2 from human, mouse, and rat.
  • Tested applications

    Suitable for: WB, IHC-P, ELISA, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic phosphopeptide derived from Human LIM Kinase around the phosphorylation site of threonine 508.

  • Positive control

    • IHC-P: Human breast carcinoma tissue slides. WB: COLO and MCF7 cell extracts.

Properties

Applications

Our Abpromise guarantee covers the use of ab38508 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/500 - 1/1000. Detects a band of approximately 65 kDa (predicted molecular weight: 73 kDa).
IHC-P 1/50 - 1/100.
ELISA 1/10000.
ICC/IF 1/100. PubMed: 19208621

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.
  • Information by UniProt
  • Database links

  • Alternative names

    • EC 2.7.1.37 antibody
    • LIM domain containing protein kinase antibody
    • LIM domain kinase 1 antibody
    • LIM kinase antibody
    • LIM motif containing protein kinase antibody
    • LIMK 1 antibody
    • LIMK antibody
    • LIMK-1 antibody
    • limk1 antibody
    • LIMK1_HUMAN antibody
    see all

Images

  • ab38508 at a 1:50 dilution staining LIM Kinase in Human breast carcinoma tissue using Immunohistochemistry, Paraffin Embedded Tissue.

    Left image : untreated.

    Right image : treated with phosphopeptide.

  • All lanes : Anti-LIM Kinase 1 (phospho T508) antibody (ab38508) at 1/500 dilution

    Lane 1 : Extract of COLO cells + peptide.
    Lane 2 : Extract of COLO cells.
    Lane 3 : Extract of MCF7 cells.

    Lysates/proteins at 30 µg per lane.

    Secondary
    All lanes : Alkaline Phosphatase AffiniPure Goat Anti-Rabbit IgG (H+L)

    Predicted band size: 73 kDa
    Observed band size: 65 kDa
    why is the actual band size different from the predicted?



    Lanes can be loaded with 5-30µg of total protein. The arrow next to the image points to LIM Kinase (phospho T508).
  • ab38508 staining LIM Kinase 1 in mouse brain tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).
    bdKO mice were anesthetized and transcardially perfused with PBS, followed by 4% paraformaldehyde (PFA) prior to brain dissection. Brains were then postfixed in 4% PFA for 2 days, embedded in paraffin and sectioned.

References

This product has been referenced in:

  • Ravindran S  et al. BDNF Induced Translation of Limk1 in Developing Neurons Regulates Dendrite Growth by Fine-Tuning Cofilin1 Activity. Front Mol Neurosci 12:64 (2019). Read more (PubMed: 30949027) »
  • Li H & Chen C Quercetin Has Antimetastatic Effects on Gastric Cancer Cells via the Interruption of uPA/uPAR Function by Modulating NF-?b, PKC-d, ERK1/2, and AMPKa. Integr Cancer Ther 17:511-523 (2018). Read more (PubMed: 28627240) »
See all 11 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Application
Western blot
Sample
Human Cultured Cells (HMC)
Gel Running Conditions
Reduced Non-Denaturing (Native) (4-20% gradient)
Loading amount
40 µg
Specification
HMC
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Oct 27 2019

Application
Western blot
Sample
Human Tissue lysate - whole (Brain)
Loading amount
20 µg
Specification
Brain
Gel Running Conditions
Reduced Denaturing
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Apr 27 2011

Answer

Thank you for your patience. I have been in contact with the originator of this antibody, and below is the detailed antigen retrieval protocol. The method of antigen retrieval was used for IHC-P as following: Deparaffinization 1. Incubate slide at 60? for 60 minutes. 2. Deparaffinize in Xylene for 10 minutes and repeat one more times. 3. Hydrate in 100% alcohol for 5 minutes, in 95% alcohol for 5 minutes, in 85% alcohol for 5 minutes, in 75% alcohol for 5 minutes. 4. Dip into Distill Water for 5 minutes. 5. Dip into TBS (50 mM Tris, 100 mM NaCl, pH 7.6), leave for 5 minutes, and repeat two times. Antigen Retrieval 6. Bring 500 - 2000 ml 10 mM citrate buffer (pH6.0) to the boil in a stainless steel pressure cooker. 7. Put the slide into staining rack and lower into pressure cooker ensuring the slide is well immersed in citrate buffer. 8. When the pressure indicator valve has risen after 3-4 minutes, incubate for 1 minute. 9. Cool the slide naturally to room temperature. 10. Dip into distilled water, leave for 5 minutes, and repeat two times. 11. Dip the slide in TBS for 5 minutes and repeat two times. 12. Immerse slides in 3% H2O2 (in fresh methanol) for 15 minutes at room temperature. 13. Wash with distilled water two times, 5 minutes each time. 14. Wash with TBS (pH 7.6) two times, 5 minutes each time. Staining with Primary Antibody 15. Dilute primary antibody with 3% BSA in TBS. Cover the tissue section on the slide with diluted primary antibody (use 50 – 150µl for each slide). 16. Incubate at 37? for 30 minutes or at room temperature for 60 minutes (The optimal incubation time, incubation temperature, and antibody dilution should be determined by the individual laboratory). 17. Wash with TBS two times, 5 minutes each time. Staining with Secondary Antibody 18. Incubate with 100-200µl Polymer Enhancer. Incubate 30 minutes at 37?. 19. Wash with TBS for 3 times, 5 minutes each time. 20. Incubate with 100-200µl Polymerized HRP and incubate 30 minutes at 37? 21. Wash with TBS for 3 times, 5 minutes each time. 22. Add DAB solution and incubate 3-10 minutes(The reaction progress and the optimal time should be determine according to microscope). 23. Wash with distilled water for 2 times, 5 minutes each time. 24. Counterstain sections in hematoxylin if required,wash with distilled water.Immerse slides in 0.1% HClethanol for 1-10 seconds, wash with distilled water. 25. Dehydrate through 95% ethanol for 1 minute, 100% ethanol for 2×3min, Xylene for 2×3min, and coverslip with mounting medium. I hope that this information helps and please do not hesitate to contact us for more information or advice in the future.

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Question
Answer

Thank you for your enquiry. From data of recent testing, this antibody recognizes both LIMK1 and LIMK2 from human, mouse, and rat. I have clarified this on the datasheet; should you have further questions please don't hesitate to contact me,

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