Recombinant Anti-Cofilin (phospho S3) antibody [EPR24753-28] - BSA and Azide free (ab283513)
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) - BSA and Azide free
- Suitable for: WB, Dot blot, IHC-P, ICC/IF, Flow Cyt (Intra)
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Cofilin (phospho S3) antibody [EPR24753-28] - BSA and Azide free
See all Cofilin-1 primary antibodies -
Description
Rabbit monoclonal [EPR24753-28] to Cofilin (phospho S3) - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: WB, Dot blot, IHC-P, 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.
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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-intra: HeLa cell. Dot Blot: Cofilin-1 (phospho S3) peptide.
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General notes
ab283513 is the carrier-free version of ab283500.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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
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
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. -
Storage buffer
pH: 7.20
Constituent: 100% PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR24753-28 -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab283513 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 |
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 18 kDa.
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Dot blot |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
Use at an assay dependent concentration.
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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Notes |
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WB
Use at an assay dependent concentration. Predicted molecular weight: 18 kDa. |
Dot blot
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
Use at an assay dependent concentration. |
Flow Cyt (Intra)
Use at an assay dependent concentration. |
Target
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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
modificationsPhosphorylated 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
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Database links
- Entrez Gene: 1072 Human
- Entrez Gene: 12631 Mouse
- Entrez Gene: 29271 Rat
- Omim: 601442 Human
- SwissProt: P23528 Human
- SwissProt: P18760 Mouse
- SwissProt: P45592 Rat
- Unigene: 170622 Human
see all -
Alternative names
- 18 kDa phosphoprotein antibody
- CFL 1 antibody
- CFL antibody
see all
Images
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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 kDaThis data was developed using ab283500, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST
Exposure time: 37 seconds
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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 kDaThis data was developed using ab283500, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST
Exposure time: 15 seconds
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All lanes : Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500) at 1/1000 dilution
Lane 1 : NIH/3T3 (mouse embryonic fibroblast), whole cell lysate (Untreated 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
Exposure time: 3 secondsThis data was developed using ab283500, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST
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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 kDaThis data was developed using ab283500, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST
Exposure time: 10 seconds
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This data was developed using ab283500, the same antibody clone in a different buffer formulation.
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
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This data was developed using ab283500, the same antibody clone in a different buffer formulation.
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
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This data was developed using ab283500, the same antibody clone in a different buffer formulation.
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
-
This data was developed using ab283500, the same antibody clone in a different buffer formulation.
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
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This data was developed using ab283500, the same antibody clone in a different buffer formulation.
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.
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This data was developed using ab283500, the same antibody clone in a different buffer formulation.
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.
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This data was developed using ab283500, the same antibody clone in a different buffer formulation.
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
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.
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (0)
ab283513 has not yet been referenced specifically in any publications.