Anti-Cofilin (phospho S3) antibody (ab100836)
Key features and details
- Rabbit polyclonal to Cofilin (phospho S3)
- Suitable for: ICC/IF, ELISA, IHC-P, WB
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-Cofilin (phospho S3) antibody
See all Cofilin primary antibodies -
Description
Rabbit polyclonal to Cofilin (phospho S3) -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, ELISA, IHC-P, WBmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide corresponding to Human Cofilin aa 1-100 (N terminal) conjugated to keyhole limpet haemocyanin.
(Peptide available asab139515) -
Positive control
- WB: Jurkat and SHSY5Y whole cell lysates. IHC-P: FFPE human breast adenocarcinoma tissue. ICC/IF: MCF7 cells.
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General notes
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab100836 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 a concentration of 5 µg/ml. ELISA Use at an assay dependent concentration. IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. WB Use a concentration of 1 µg/ml. Detects a band of approximately 20 kDa (predicted molecular weight: 18 kDa). 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
- Omim: 601442 Human
- SwissProt: P23528 Human
- Unigene: 170622 Human
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Alternative names
- 18 kDa phosphoprotein antibody
- CFL 1 antibody
- CFL antibody
see all
Images
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Western blot - Anti-Cofilin (phospho S3) antibody (ab100836)All lanes : Anti-Cofilin (phospho S3) antibody (ab100836) at 1 µg/ml
Lane 1 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lane 2 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate
Lane 3 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate with Immunizing peptide at 1 µg/ml
Lane 4 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate with Immunizing peptide at 1 µg/ml
Lane 5 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate with Non modified peptide at 1 µg/ml
Lane 6 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate with Non modified peptide at 1 µg/ml
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 18 kDa
Observed band size: 20 kDa why is the actual band size different from the predicted?
Additional bands at: 110 kDa, 150 kDa, 33 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 180 seconds -
Immunocytochemistry/ Immunofluorescence - Anti-Cofilin (phospho S3) antibody (ab100836)ICC/IF image of ab100836 stained MCF7 cells. The cells were 4% formaldehyde 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 (ab100836, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 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. This antibody also gave a positive result in 4% formaldehyde fixed (10 min) HeLa cells at 5µg/ml,.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cofilin (phospho S3) antibody (ab100836)IHC image of ab100836 staining in human breast adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica 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 ab100836, 1µg/ml, for 15 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.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times. -
ELISA - Anti-Cofilin (phospho S3) antibody (ab100836)ab100836 was tested using an Indirect ELISA approach. The wells were coated with peptide (1µg/ml at 100µl/well) overnight at 4°C, followed by a 5% BSA blocking step for 1 hour at room temperature. The primary Ab was then added at a dilution range of 1- 0.00025µg/ml (100µl/well) for 1hr at room temperature. A HRP-conjugated anti-rabbit IgG (heavy and light chain) was used as a secondary antibody at 1:20,000 dilution for 1hr at room temperature.
Protocols
Datasheets and documents
References (2)
ab100836 has been referenced in 2 publications.
- Skaria T et al. Wnt5A/Ryk signaling critically affects barrier function in human vascular endothelial cells. Cell Adh Migr 11:24-38 (2017). PubMed: 27159116
- Skaria T et al. IL-4 Causes Hyperpermeability of Vascular Endothelial Cells through Wnt5A Signaling. PLoS One 11:e0156002 (2016). WB, IF . PubMed: 27214384
Images
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Western blot - Anti-Cofilin (phospho S3) antibody (ab100836)All lanes : Anti-Cofilin (phospho S3) antibody (ab100836) at 1 µg/ml
Lane 1 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lane 2 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate
Lane 3 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate with Immunizing peptide at 1 µg/ml
Lane 4 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate with Immunizing peptide at 1 µg/ml
Lane 5 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate with Non modified peptide at 1 µg/ml
Lane 6 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate with Non modified peptide at 1 µg/ml
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 18 kDa
Observed band size: 20 kDa why is the actual band size different from the predicted?
Additional bands at: 110 kDa, 150 kDa, 33 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 180 seconds
-
Immunocytochemistry/ Immunofluorescence - Anti-Cofilin (phospho S3) antibody (ab100836)
ICC/IF image of ab100836 stained MCF7 cells. The cells were 4% formaldehyde 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 (ab100836, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 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. This antibody also gave a positive result in 4% formaldehyde fixed (10 min) HeLa cells at 5µg/ml,.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cofilin (phospho S3) antibody (ab100836)IHC image of ab100836 staining in human breast adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica 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 ab100836, 1µg/ml, for 15 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.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times. -
ELISA - Anti-Cofilin (phospho S3) antibody (ab100836)
ab100836 was tested using an Indirect ELISA approach. The wells were coated with peptide (1µg/ml at 100µl/well) overnight at 4°C, followed by a 5% BSA blocking step for 1 hour at room temperature. The primary Ab was then added at a dilution range of 1- 0.00025µg/ml (100µl/well) for 1hr at room temperature. A HRP-conjugated anti-rabbit IgG (heavy and light chain) was used as a secondary antibody at 1:20,000 dilution for 1hr at room temperature.
