Anti-Cofilin (phospho S3) antibody (ab12866)

Overview

  • Product nameAnti-Cofilin (phospho S3) antibody
    See all Cofilin primary antibodies
  • Description
    Rabbit polyclonal to Cofilin (phospho S3)
  • Tested applicationsSuitable for: ICC/IF, WB, IHC-FoFr, IHC-Pmore details
  • Species reactivity
    Reacts with: Rat, Dog, Human
    Predicted to work with: Mouse
  • Immunogen

    Synthetic peptide derived from a region of human cofilin that contains serine 3. The sequence is conserved in mouse and rat.

  • Positive control
    • MDCK cells treated with staurosporine; Jurkat cells treated with hydrogen peroxide.

Properties

Applications

Our Abpromise guarantee covers the use of ab12866 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
ICC/IF Use at an assay dependent concentration.
WB 1/1000. Detects a band of approximately 20 kDa.
IHC-FoFr Use at an assay dependent concentration. PubMed: 17652593
IHC-P Use at an assay dependent concentration. PubMed: 20797537

Target

  • FunctionControls reversibly actin polymerization and depolymerization in a pH-sensitive manner. It has the ability to bind G- and F-actin in a 1:1 ratio of cofilin to actin. It is the major component of intranuclear and cytoplasmic actin rods.
  • Tissue specificityWidely distributed in various tissues.
  • Sequence similaritiesBelongs to the actin-binding proteins ADF family.
    Contains 1 ADF-H domain.
  • Post-translational
    modifications
    Phosphorylated on Ser-3 in resting cells.
  • Cellular localizationNucleus matrix. Cytoplasm > cytoskeleton. Almost completely in nucleus in cells exposed to heat shock or 10% dimethyl sulfoxide.
  • Information by UniProt
  • Database links
  • Alternative names
    • 18 kDa phosphoprotein antibody
    • CFL 1 antibody
    • CFL antibody
    • CFL1 antibody
    • COF1_HUMAN antibody
    • Cofilin 1 antibody
    • Cofilin 1 non muscle antibody
    • Cofilin antibody
    • Cofilin non muscle isoform antibody
    • Cofilin-1 antibody
    • epididymis secretory protein Li 15 antibody
    • HEL-S-15 antibody
    • non-muscle isoform antibody
    • p18 antibody
    see all

Anti-Cofilin (phospho S3) antibody images

  • Peptide Competition and Phosphatase Treatment Lysates prepared from MDCK cells treated with staurosporine (1) or left untreated (2-6) were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were either left untreated (1-5) or treated with lambda phosphatase (6), blocked with a 5% BSA-TBST buffer for one hour at room temperature, and incubated with the ab12866 antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1, 2, 6), the non phosphopeptide corresponding to the immunogen (3), a generic phosphoserine-containing peptide (4) or, the phosphopeptide immunogen (5). After washing, membranes were incubated with goat F(ab)2 anti-rabbit IgG HRP conjugate and bands were detected using the Pierce SuperSignal method. The data show that only the peptide corresponding to cofilin [pS3] blocks the antibody signal. The data also show that phosphatase stripping eliminates the signal, verifying that the antibody is phospho-specific.
  • ICC/IF image of ab12866 stained MCF7 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 non-specific protein-protein interactions. The cells were then incubated with the antibody (ab12866, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 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.

References for Anti-Cofilin (phospho S3) antibody (ab12866)

This product has been referenced in:
  • Vajda F  et al. Cell type-specific Nogo-A gene ablation promotes axonal regeneration in the injured adult optic nerve. Cell Death Differ 22:323-35 (2015). Mouse . Read more (PubMed: 25257170) »
  • Speranza L  et al. Activation of 5-HT7 receptor stimulates neurite elongation through mTOR, Cdc42 and actin filaments dynamics. Front Behav Neurosci 9:62 (2015). WB ; Mouse . Read more (PubMed: 25814944) »

See all 22 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (Primary cortical neuronal cells)
Permeabilization Yes - 0.2% Triton X-100
Specification Primary cortical neuronal cells
Blocking step casein solution + Tween 20 as blocking agent for 20 minute(s) · Concentration: 0.2% · Temperature: 25°C
Fixative Ethanol
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Verified customer

Submitted Aug 18 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - whole (Nervous Tissue (Nucleus Accumbens))
Gel Running Conditions Reduced Denaturing (10% gel)
Loading amount 40 µg
Specification Nervous Tissue (Nucleus Accumbens)
Blocking step Li-Cor Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 50% · Temperature: 21°C
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Mr. Jeffrey Lenz

Verified customer

Submitted Jan 27 2016

Application Western blot
Loading amount 75 µg
Gel Running Conditions Reduced Denaturing (12%)
Sample Mouse Tissue lysate - whole (brain tissue)
Specification brain tissue
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Nov 10 2014

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing (12)
Sample Human Tissue lysate - whole (Cerebellum, Brain)
Specification Cerebellum, Brain
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Apr 02 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Loading amount 50 µg
Gel Running Conditions Reduced Denaturing
Sample Rat Tissue lysate - whole (Brain)
Specification Brain
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
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Submitted Jun 24 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Loading amount 50 µg
Gel Running Conditions Reduced Denaturing (12%)
Sample Human Cell lysate - whole cell (Neuroblastoma)
Specification Neuroblastoma
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
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Submitted Jun 24 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Neuroblastoma)
Specification Neuroblastoma
Fixative Formaldehyde
Permeabilization Yes - 0.1% Trition X
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 21°C
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Submitted May 13 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Neuroblastoma)
Specification Neuroblastoma
Fixative Paraformaldehyde
Permeabilization Yes - 0.01% Triton X
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 20°C
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Submitted Sep 04 2012

Thank you very much for your call today and for your questions about ab12866.

I've further discussed the antibody and proposed staining method with my colleague, and even though we have not tested the antibody in exactly this way,your protoco...

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Thank you for your inquiry.
That antibody is our ab12866, rabbit polyclonal to Cofilin (phospho S3).
http://www.abcam.com/Cofilin-phospho-S3-antibody-ab12866.html
We have that reference listed on our publications page.
http://www.ab...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"