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RecombinantRabMAb

Recombinant Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500)

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Western blot - Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500)
  • Western blot - Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500)
  • Western blot - Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500)
  • Western blot - Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500)
  • Immunocytochemistry/ Immunofluorescence - Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500)
  • Flow Cytometry (Intracellular) - Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500)
  • Dot Blot - Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500)
  • Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR24753-28] to Cofilin (phospho S3)
  • Suitable for: WB, IHC-P, Dot blot, ICC/IF, Flow Cyt (Intra)
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-Cofilin (phospho S3) antibody [EPR24753-28]
    See all Cofilin-1 primary antibodies

  • Description

    Rabbit monoclonal [EPR24753-28] to Cofilin (phospho S3)

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, Dot blot, ICC/IF, Flow Cyt (Intra)more details
    Unsuitable for: IP

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HeLa, 293T, NIH/3T3 and C6 lysates all without alkaline phosphatase treatment. IHC-P: Human breast carcinoma, Human stomach, Mouse colon, Rat colon tissues all without alkaline phosphatase treatment. ICC/IF: HeLa cells without alkaline phosphatase treatment. Flow Cyt: HeLa cell. Dot Blot: Cofilin-1 (phospho S3) peptide.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab283500 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
1/1000. Predicted molecular weight: 18 kDa.
IHC-P
1/5000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Dot blot
1/1000.
ICC/IF
1/1000.
Flow Cyt (Intra)
1/600.
Notes
WB
1/1000. Predicted molecular weight: 18 kDa.
IHC-P
1/5000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Dot blot
1/1000.
ICC/IF
1/1000.
Flow Cyt (Intra)
1/600.
Application notes
Is unsuitable for IP.

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

    see all

  • 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

  • Western blot - Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500)
    Western blot - Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500)
    All lanes : Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500) at 1/1000 dilution

    Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate (Untreated membrane)
    Lane 2 : HeLa(human cervix adenocarcinoma epithelial cell), whole cell lysate (Phosphatase treated membrane)

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 18 kDa
    Observed band size: 18 kDa



    Blocking and diluting buffer and concentration: 5% NFDM/TBST

    Exposure time: 37 seconds

     

  • Western blot - Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500)
    Western blot - Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500)
    All lanes : Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500) at 1/1000 dilution

    Lane 1 : 293T (human embryonic kidney epithelial cell), whole cell lysate (Untreated membrane)
    Lane 2 : 293T (Phosphatase treated membrane)

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/50000 dilution

    Predicted band size: 18 kDa
    Observed band size: 18 kDa



    Blocking and diluting buffer and concentration: 5% NFDM/TBST

    Exposure time: 15 seconds

     

  • Western blot - Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500)
    Western blot - Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500)
    All lanes : Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500) at 1/1000 dilution

    Lane 1 : 293T (Phosphatase treated membrane)
    Lane 2 : NIH/3T3 (Phosphatase treated membrane)

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/50000 dilution

    Predicted band size: 18 kDa
    Observed band size: 18 kDa



    Blocking and diluting buffer and concentration: 5% NFDM/TBST

    Exposure time: 3 seconds

     

  • Western blot - Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500)
    Western blot - Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500)
    All lanes : Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500) at 1/1000 dilution

    Lane 1 : C6 (rat glial tumor glial cell), whole cell lysate (Untreated membrane)
    Lane 2 : C6 (Phosphatase treated membrane)

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/50000 dilution

    Predicted band size: 18 kDa
    Observed band size: 18 kDa



    Blocking and diluting buffer and concentration: 5% NFDM/TBST

    Exposure time: 10 seconds

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500)

    Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labelling Cofilin (phospho S3) with ab283500 at 1/5000 (0.121 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human breast carcinoma without alkaline phosphatase treatment (image A). (PMID: 27799793) No signal was detected when tissues were treated with alkaline phosphatase (image B). The section was incubated with ab283500 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500)

    Immunohistochemical analysis of paraffin-embedded Human stomach tissue labelling Cofilin (phospho S3) with ab283500 at 1/5000 (0.121 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human stomach without alkaline phosphatase treatment (image A). No signal was detected when tissues were treated with alkaline phosphatase (image B). The section was incubated with ab283500 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500)

    Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labelling Cofilin (phospho S3) with ab283500 at 1/5000 (0.121 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse colon without alkaline phosphatase treatment (image A). (PMID: 15649475) No signal was detected when tissues were treated with alkaline phosphatase (image B). The section was incubated with ab283500 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500)

    Immunohistochemical analysis of paraffin-embedded Rat colon tissue labelling Cofilin (phospho S3) with ab283500 at 1/5000 (0.121 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on rat colon without alkaline phosphatase treatment (image A). No signal was detected when tissues were treated with alkaline phosphatase (image B). The section was incubated with ab283500 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

  • Immunocytochemistry/ Immunofluorescence - Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500)
    Immunocytochemistry/ Immunofluorescence - Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa cells labelling Cofilin (phospho S3) with ab283500 at 1/1000 (0.604 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing nuclear and cytoplasmic staining in HeLa cells, no staining in HeLa treated with Alkaline Phosphatase (37°C) for 1 hour is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

  • Flow Cytometry (Intracellular) - Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500)
    Flow Cytometry (Intracellular) - Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500)

    Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling Cofilin (phospho S3) with ab283500 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

  • Dot Blot - Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500)
    Dot Blot - Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500)

    Dot blot analysis of Cofilin (phospho S3) using ab283500 at 1/1000 (0.604 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100,000 dilution.

    Lane 1: Cofilin-1 (phospho S3) peptide

    Lane 2: Cofilin-1 non-phospho peptide

    Exposure time: 3 minutes

    Blocking and diluting buffer and concentration: 5% NFDM/TBST

  • Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500)
    Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (0)

Publishing research using ab283500? Please let us know so that we can cite the reference in this datasheet.

ab283500 has not yet been referenced specifically in any publications.

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