Overview

  • Product name
  • Description
    Rabbit polyclonal to Cofilin
  • Tested applications
    Suitable for: IHC-FoFr, ICC/IF, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Sheep, Cow, Pig
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 150 to the C-terminus of Human Cofilin.

    (Peptide available as ab42823.)

  • Positive control
    • WB: HeLa, Jurkat, A431, HEK293, MCF7, SHSY-5Y, PC12 and NIH 3T3 whole cell lysates. ICC/IF: HeLa cells. IHC-P: Human breast carcinoma tissue.

Properties

Applications

Our Abpromise guarantee covers the use of ab42824 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
IHC-FoFr 1/2000 - 1/3000.
ICC/IF Use a concentration of 1 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 20 kDa (predicted molecular weight: 19 kDa).
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Target

  • Function
    Controls 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 specificity
    Widely distributed in various tissues.
  • Sequence similarities
    Belongs to the actin-binding proteins ADF family.
    Contains 1 ADF-H domain.
  • Post-translational
    modifications
    Phosphorylated on Ser-3 in resting cells.
  • Cellular localization
    Nucleus 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

Images

  • All lanes : Anti-Cofilin antibody (ab42824) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 4 : HEK293 Human embryonic kidney cell line Whole Cell Lysate
    Lane 5 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
    Lane 6 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 19 kDa
    Observed band size : 20 kDa (why is the actual band size different from the predicted?)
  • ICC/IF image of ab42824 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab42824, 1µg/ml) for 1h at room temperature. 1% BSA / 10% normal serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. 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). DAPI was used to stain the cell nuclei (blue).

  • IHC image of Cofilin staining in human breast carcinoma FFPE section, performed on a 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 ab42824, 1µg/ml, for 8 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
  • Anti-Cofilin antibody (ab42824) at 1 µg/ml + Recombinant Human Cofilin protein (ab62958) at 0.01 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Predicted band size : 19 kDa
  • All lanes : Anti-Cofilin antibody (ab42824) at 1 µg/ml

    Lane 1 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
    Lane 2 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 19 kDa
    Observed band size : 20 kDa (why is the actual band size different from the predicted?)


    Exposure time : 30 seconds

    This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab42824 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

References

This product has been referenced in:
  • Lee HJ  et al. Fluid shear stress activates YAP1 to promote cancer cell motility. Nat Commun 8:14122 (2017). WB ; Human . Read more (PubMed: 28098159) »
  • Rao AN  et al. Pharmacologically increasing microtubule acetylation corrects stress-exacerbated effects of organophosphates on neurons. Traffic 18:433-441 (2017). WB ; Rat . Read more (PubMed: 28471062) »

See all 20 Publications for this product

Customer reviews and Q&As

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Pig Cell lysate - whole cell (isolated hepatocyte cells)
Gel Running Conditions
Reduced Denaturing (4-12% gradient gel)
Loading amount
10 µg
Specification
isolated hepatocyte cells
Blocking step
BSA as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 3% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted May 08 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Pig Tissue sections (endometrium)
Antigen retrieval step
Heat mediated
Permeabilization
No
Specification
endometrium
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Feb 12 2016

Application
Western blot
Sample
Pig Tissue lysate - whole (endometrial tissue lysate)
Gel Running Conditions
Reduced Denaturing (15%)
Loading amount
10 µg
Specification
endometrial tissue lysate
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted Feb 11 2016

Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from ab54532. The details you have kindly provided will provide us with vital information for our monitoring of product quality
...

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Thank you for contacting us.

ab42824 is tested in western blot with human and rat samples and is fully guaranteed (http://www.abcam.com/index.html?pageconfig=abpromise). The observed ˜30kDa and ˜90kDa bands might show slight non specificity of...

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Thank you for your reply.

Neither ab42824 or ab12866 show any cross reactivity with either Destrin (ADF) or phosphor Destrin. As neither of these antibodies has been shown to work on monkey tissue then they are both eligible for our testing d...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Rat Tissue sections (brain sections)
Specification
brain sections
Fixative
Paraformaldehyde
Antigen retrieval step
None
Permeabilization
No
Username

Dr. Sophie Pezet

Verified customer

Submitted Sep 25 2008

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

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