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

Rabbit Recombinant Monoclonal Cofilin-1 phospho S3 antibody. Suitable for ICC/IF, Dot, WB, Flow Cyt (Intra), IHC-P and reacts with Human, Rat, Synthetic peptide, Mouse samples. Cited in 1 publication.

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Images

Western blot - Anti-Cofilin (phospho S3) antibody [EPR24753-28] (AB283500), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	ICC/IF	IP	Dot	WB	Flow Cyt (Intra)	IHC-P
Human	Tested	Not recommended	Expected	Tested	Tested	Tested
Mouse	Not recommended	Not recommended	Expected	Tested	Expected	Tested
Rat	Expected	Not recommended	Expected	Tested	Expected	Tested
Synthetic peptide	Not recommended	Not recommended	Tested	Not recommended	Not recommended	Not recommended

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/1000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Synthetic peptide	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info -	Notes -
Species Synthetic peptide	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Synthetic peptide	Dilution info 1/1000	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Human, Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000	Notes -
Species Rat	Dilution info 1/1000	Notes -
Species Human	Dilution info 1/1000	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Synthetic peptide	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/600	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse	Dilution info 1/600	Notes -
Species Rat	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Synthetic peptide	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/5000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/5000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info 1/5000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species	Dilution info	Notes
Species Synthetic peptide	Dilution info -	Notes -

Target data

See full target information CFL1 phospho S3

Function

Binds to F-actin and exhibits pH-sensitive F-actin depolymerizing activity (PubMed:11812157). In conjunction with the subcortical maternal complex (SCMC), plays an essential role for zygotes to progress beyond the first embryonic cell divisions via regulation of actin dynamics (PubMed:15580268). Required for the centralization of the mitotic spindle and symmetric division of zygotes (By similarity). Plays a role in the regulation of cell morphology and cytoskeletal organization in epithelial cells (PubMed:21834987). Required for the up-regulation of atypical chemokine receptor ACKR2 from endosomal compartment to cell membrane, increasing its efficiency in chemokine uptake and degradation (PubMed:23633677). Required for neural tube morphogenesis and neural crest cell migration (By similarity).

Alternative names

CFL, CFL1, Cofilin-1, 18 kDa phosphoprotein, p18

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR24753-28
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

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

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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Cofilin-1 also known just as cofilin is a small actin-modulating protein with a molecular weight of approximately 19 kDa. It is part of the actin chemical structure regulation and plays an important role in actin filament dynamics by severing and depolymerizing actin filaments. Cofilin-1 expresses widely in both muscle and non-muscle tissues modulating the actin cytoskeleton which is critical for cell movement shape and various signaling pathways. The phosphorylated form of this protein known as phospho-cofilin or p-cofilin is another significant state to consider.

Biological function summary

Through its interaction with actin filaments cofilin-1 functions to regulate the length and turnover of the actin cytoskeleton within cells. Cofilin-1 is part of a critical complex where it binds to actin monomers and filaments to facilitate the disassembly and recycling of actin. This regulation impacts cellular activities such as migration endocytosis and division influencing how cells respond to internal and external signals. Cofilin-1's ability to bind and sever actin filaments forms an important part of its functional mechanism altering the cytoskeletal architecture for various cellular processes.

Pathways

Cofilin-1 participates principally in the actin dynamics component of the cytoskeleton remodeling pathway. Within this framework cofilin-1 acts alongside other proteins like actin-depolymerizing factor (ADF) and tropomyosin influencing assembly and disassembly cycles of actin filaments. The Rho family of GTPases regulates cofilin-1 activity through signaling pathways such as the Rho/ROCK/LIMK pathway. This signaling regulation affects actin filament organization impacting cellular processes like axonal guidance and directed cell movement.

Associated diseases and disorders

There is a strong connection between cofilin-1 dysregulation and neurodegenerative diseases such as Alzheimer's disease and certain types of cancer. In Alzheimer's disease cofilin-1 activity influenced by abnormal cofilin phosphorylation forms part of the pathological mechanisms that lead to synaptic dysfunction and neurofibrillary tangles. In the context of cancer cofilin-1 collaborates with proteins such as RhoA and LIM kinase contributing to cancer cell invasion and metastasis by reorganizing the actin cytoskeleton to facilitate cell motility. Understanding these interactions helps in targeting therapeutic strategies against these conditions.

Product promise

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11 product images

Western blot - Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500)
Blocking and diluting buffer and concentration: 5% NFDM/TBST
Exposure time: 37 seconds
All lanes: Western blot - 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) at 10 µg
Lane 2: HeLa(human cervix adenocarcinoma epithelial cell), whole cell lysate (Phosphatase treated membrane) at 10 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution
Predicted band size: 18 kDa
Observed band size: 18 kDa
Western blot - Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500)
Blocking and diluting buffer and concentration: 5% NFDM/TBST
All lanes: Western blot - Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500) at 1/1000 dilution
Lane 1: NIH/3T3 (mouse embryonic fibroblast), whole cell lysate (Untreated membrane) at 10 µg
Lane 2: NIH/3T3 (Phosphatase treated membrane) at 10 µg
Secondary
All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution
Predicted band size: 18 kDa
Observed band size: 18 kDa
Exposure time: 3s
Western blot - Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500)
Blocking and diluting buffer and concentration: 5% NFDM/TBST
Exposure time: 3 seconds
All lanes: Western blot - Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500) at 1/1000 dilution
Lane 1: 293T (Phosphatase treated membrane) at 10 µg
Lane 2: NIH/3T3 (Phosphatase treated membrane) at 10 µg
Secondary
All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution
Predicted band size: 18 kDa
Observed band size: 18 kDa
Western blot - Anti-Cofilin (phospho S3) antibody [EPR24753-28] (ab283500)
Blocking and diluting buffer and concentration: 5% NFDM/TBST
Exposure time: 10 seconds
All lanes: Western blot - 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) at 10 µg
Lane 2: C6 (Phosphatase treated membrane) at 10 µg
Secondary
All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution
Predicted band size: 18 kDa
Observed band size: 18 kDa
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 (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 (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)
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 (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 (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)
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 (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 (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)
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)
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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed 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. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed 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 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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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, Goat Anti-Rabbit IgG H&L (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 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 (Goat Anti-Rabbit IgG H&L (HRP) 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