Name: Anti-S100A4 antibody [EPR14639(2)]
SKU: ab197896
Rating: 4 (8 reviews)

Anti-S100A4 antibody [EPR14639(2)] (ab197896) is a rabbit monoclonal antibody detecting S100A4 in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IP, IHC-P, ICC/IF. Suitable for Human, Mouse, Rat.

- Clone EPR14639(2) is the most cited clone to S100A4

- KO validated for confirmed specificity

- Biophysical QC for unrivalled batch-batch consistency

- Over 60 publications

Images

Western blot - Anti-S100A4 antibody [EPR14639(2)] (AB197896), 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.

Abcam Recommends

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

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

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/40	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 Mouse	Dilution info Use at an assay dependent concentration.	Notes -

Predicted
Predicted

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes -

Tested
Tested

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

Predicted
Predicted

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/250	Notes -

Expected
Expected

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

Predicted
Predicted

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/250	Notes -

Expected
Expected

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

Predicted
Predicted

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/2000	Notes We do not guarantee IHC-P for mouse and rat. 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 Rat	Dilution info -	Notes We do not guarantee IHC-P for mouse and rat. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Mouse	Dilution info -	Notes We do not guarantee IHC-P for mouse and rat. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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Target data

See full target information S100A4

Function

Calcium-binding protein that plays a role in various cellular processes including motility, angiogenesis, cell differentiation, apoptosis, and autophagy (PubMed:16707441, PubMed:23752197, PubMed:30713770). Increases cell motility and invasiveness by interacting with non-muscle myosin heavy chain (NMMHC) IIA/MYH9 (PubMed:16707441). Mechanistically, promotes filament depolymerization and increases the amount of soluble myosin-IIA, resulting in the formation of stable protrusions facilitating chemotaxis (By similarity). Modulates also the pro-apoptotic function of TP53 by binding to its C-terminal transactivation domain within the nucleus and reducing its protein levels (PubMed:23752197). Within the extracellular space, stimulates cytokine production including granulocyte colony-stimulating factor and CCL24 from T-lymphocytes (By similarity). In addition, stimulates T-lymphocyte chemotaxis by acting as a chemoattractant complex with PGLYRP1 that promotes lymphocyte migration via CCR5 and CXCR3 receptors (PubMed:26654597, PubMed:30713770).

Alternative names

CAPL, MTS1, S100A4, Protein S100-A4, Calvasculin, Metastasin, Placental calcium-binding protein, Protein Mts1, S100 calcium-binding protein A4

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: EPR14639(2)
Purification technique: Affinity purification Protein A
Specificity: Based on sequence homologies, the antibody may cross-react with other proteins of the same family (S100A1-12). We did not perform any experiments to confirm this.
We do not guarantee IHC-P for mouse and rat. Some optimisation may be required for detection of the target protein due to low levels of endogenous expression in some samples. Please see images below for suitable positive controls.
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

Product Specifications
Anti-S100A4 antibody [EPR14639(2)] (ab197896) was developed by Abcam using patented rabbit monoclonal antibody technology and is validated for use in Flow Cyt (Intra), ICC/IF, IHC-P, IP, WB in human, mouse, rat samples.
Anti-S100A4 antibody [EPR14639(2)] (ab197896) specifically detects S100A4 (UniProt ID: P26447; Molecular weight: 12kDa) and is sold in 100 µL and 1 mL selling sizes.

Quality and Validation
Abcam's high quality manufacturing and validation processes ensure Anti-S100A4 antibody [EPR14639(2)] (ab197896) has high sensitivity and specificity alongside high lot-to-lot consistency and reproducibility.
The specificity of Anti-S100A4 antibody [EPR14639(2)] (ab197896) has been confirmed by testing in knockout samples.
Anti-S100A4 antibody [EPR14639(2)] (ab197896) has been cited over 62 times in peer reviewed journals and is trusted by the scientific community.
Anti-S100A4 antibody [EPR14639(2)] (ab197896) has 8 independent reviews from customers.

Related Products
Antibody clone EPR14639(2) is also available pre-conjugated to a variety of labels for your convenience - Alexa Fluor® 488 (Alexa Fluor® 488 Anti-S100A4 antibody [EPR14639(2)] ab208566).

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

What are the advantages of a recombinant monoclonal antibody?
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.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

12 product images

Western blot - Anti-S100A4 antibody [EPR14639(2)] (ab197896)
False colour image of Western blot: Anti-S100A4 antibody [EPR14639(2)] staining at 1/1000 dilution, shown in green;loading control Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) staining at 1/20000 dilution, shown in red. In Western blot, ab197896 was shown to bind specifically to S100A4. A band was observed at 11 kDa in wild-type HeLa and A549 cell lysates with no signal observed at this size in S100A4 knockout HeLa cell line Human S100A4 knockout HeLa cell line ab265709 (knockout cell lysate Human S100A4 knockout HeLa cell lysate ab257046) and S100A4 knockout A549 cell line Human S100A4 knockout A549 cell line ab261865 (knockout cell lysate Human S100A4 knockout A549 cell lysate ab261674). To generate this image, wild-type and S100A4 knockout HeLa and S100A4 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.
All lanes: Western blot - Anti-S100A4 antibody [EPR14639(2)] (ab197896) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: S100A4 knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human S100A4 knockout HeLa cell line (Human S100A4 knockout HeLa cell line ab265709)
Lane 3: Wild-type A549 cell lysate at 20 µg
Lane 4: S100A4 knockout A549 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 12 kDa
Observed band size: 11 kDa
Western blot - Anti-S100A4 antibody [EPR14639(2)] (ab197896)
Lanes 1 - 4: Merged signal (red and green). Green - ab197896 observed at 12 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.
ab197896 was shown to recognize S100A4 in wild-type A549 cells as signal was lost at the expected MW in S100A4 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and S100A4 knockout samples were subjected to SDS-PAGE. ab197896 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-S100A4 antibody [EPR14639(2)] (ab197896) at 1/1000 dilution
Lane 1: Wild-type A549 whole cell lysate at 20 µg
Lane 2: S100A4 knockout A549 whole cell lysate at 20 µg
Lane 3: A375 whole cell lysate at 20 µg
Lane 4: HeLa whole cell lysate at 20 µg
Predicted band size: 12 kDa
Observed band size: 12 kDa
Western blot - Anti-S100A4 antibody [EPR14639(2)] (ab197896)
Lanes 1 - 2: Merged signal (red and green). Green - ab197896 observed at 11 kDa. Red - loading control Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
ab197896 was shown to react with S100A4 in wild-type HeLa cells in Western blot with loss of signal observed in S100A4 knockout cell line Human S100A4 knockout HeLa cell line ab265709 (S100A4 knockout cell lysate Human S100A4 knockout HeLa cell lysate ab257046). Wild-type HeLa and S100A4 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab197896 and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
All lanes: Western blot - Anti-S100A4 antibody [EPR14639(2)] (ab197896) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: S100A4 knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human S100A4 knockout HeLa cell line (Human S100A4 knockout HeLa cell line ab265709)
Performed under reducing conditions.
Predicted band size: 12 kDa
Observed band size: 11 kDa
Western blot - Anti-S100A4 antibody [EPR14639(2)] (ab197896)
Lanes 1 - 2: Merged signal (red and green). Green - ab197896 observed at 12 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 37 kDa.
ab197896 Recombinant Anti-S100A4 antibody [EPR14639(2)] was shown to specifically react with S100A4 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261758 (knockout cell lysate ab257045) was used. Wild-type and S100A4 knockout samples were subjected to SDS-PAGE. ab197896 and Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (Anti-alpha Tubulin antibody [EP1332Y] - Loading Control ab52866) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-S100A4 antibody [EPR14639(2)] (ab197896) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: S100A4 knockout HeLa cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 12 kDa
Observed band size: 12 kDa
Immunocytochemistry/ Immunofluorescence - Anti-S100A4 antibody [EPR14639(2)] (ab197896)
Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling S100A4 with ab197896 at 1/200 (10 μg/mL). Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% TritonX-100. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077, AlexaFluor^®488 Goat anti-Rabbit at 1/1000 (2 μg/mL) was used as the secondary antibody. Cells were counterstained with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889, Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at 1/200 (2.5 μg/mL). Nuclear counter stain was DAPI (blue). Confocal image showing positive staining in HeLa cell line.
Immunoprecipitation - Anti-S100A4 antibody [EPR14639(2)] (ab197896)
ab197896 at 1/40 immunoprecipitating S100A4 in A549 whole cell lysate observed at 12 KDa.
Lane 1 (input): A549 whole cell lysate 10μg
Lane 2 (+): ab197896 + A549 whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab197896 in A549 whole cell lysate
For western blotting, Panel A: ab197896, 1:1000; Panel B: Anti-S100A4 antibody [EPR2761(2)] ab124805, 1:1000 and anti-rabbit IgG (HRP), specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).
Blocking/Diluting buffer and concentration: 5% NFDM/TBST.
All lanes: Immunoprecipitation - Anti-S100A4 antibody [EPR14639(2)] (ab197896)
Predicted band size: 12 kDa
Exposure time: 10s
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100A4 antibody [EPR14639(2)] (ab197896)
Immunohistochemical analysis of paraffin-embedded Human gastric carcinoma tissue labeling S100A4 using ab197896 at 1/2000 dilution. Goat Anti-Rabbit IgG H&L (HRP) Goat Anti-Rabbit IgG H&L (HRP) ab97051 was used as a secondary antibody at 1/500 dilution. Cells were counterstained with Hematoxylin.
Inset image: negative control obtained using PBS instead of ab197896 and secondary antibody only.
Note: Cytoplasm and nuclear staining on human gastric carcinoma tissue was observed.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cytometry (Intracellular) - Anti-S100A4 antibody [EPR14639(2)] (ab197896)
Intracellular Flow Cytometry analysis of Jurkat cells labelling S100A4 with ab197896 at 1/250 (red). Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. An Alexa Fluorr^® 488-conjugated goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100A4 antibody [EPR14639(2)] (ab197896)
Immunohistochemical analysis of paraffin-embedded Human cervix carcinoma tissue labeling S100A4 using ab197896 at 1/2000 dilution. Goat Anti-Rabbit IgG H&L (HRP) Goat Anti-Rabbit IgG H&L (HRP) ab97051 was used ay 1/500 dilution as a secondary antibody and cells were counterstained with Hematoxylin.
Inset image: negative control obtained using PBS instead of ab197896 and secondary antibody only.
Note: Nuclear and cytoplasm staining on cervix carcinoma tissue was observed.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100A4 antibody [EPR14639(2)] (ab197896)
Immunohistochemical analysis of paraffin-embedded Human lung carcinoma tissue labeling S100A4 using ab197896 at 1/2000 dilution. Goat Anti-Rabbit IgG H&L (HRP) Goat Anti-Rabbit IgG H&L (HRP) ab97051 was used as a secondary antibody at a dilution of 1/500 and cells were counterstained with Hematoxylin.
Inset image: negative control obtained using PBS instead of ab197896 and secondary antibody only.
Note: Nuclear and weakly staining on lung carcinoma tissue was observed.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-S100A4 antibody [EPR14639(2)] (ab197896)
Blocking and dilution buffer: 5% NFDM/TBST.
Some optimisation may be required for detection of the target protein due to low levels of endogenous expression in some samples.
All lanes: Western blot - Anti-S100A4 antibody [EPR14639(2)] (ab197896) at 1/1000 dilution
Lane 1: A549 (Human lung carcinoma epithelial cell) whole cell lysates at 20 µg
Lane 2: NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates at 20 µg
Lane 3: Raw264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates at 20 µg
Lane 4: Mouse spleen tissue lysates at 20 µg
Lane 5: Mouse bone marrow tissue lysates at 20 µg
Lane 6: Mouse heart tissue lysates at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 12 kDa
Observed band size: 12 kDa
Exposure time: 44s
Western blot - Anti-S100A4 antibody [EPR14639(2)] (ab197896)
Blocking and diluting buffer and concentration: 5% NFDM /TBST.
Compared with ab197896, Anti-S100A4 antibody [EPR2761(2)] ab124805 has higher sensitivity. We recommend Anti-S100A4 antibody [EPR2761(2)] ab124805 as an alternative for testing western blot.
We suggest optimizing experimental protocols (increasing lysate amount, using lower dilution or higher sensitivity ECL substrate) to improve results when using ab197896 in western blot.
All lanes: Western blot - Anti-S100A4 antibody [EPR14639(2)] (ab197896) at 1/1000 dilution
Lane 1: Human tonsil tissue lysate at 20 µg
Lane 2: A549 (Human lung carcinoma epithelial cell) whole cell lysates at 20 µg
Lane 3: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Observed band size: 12 kDa
Exposure time: 10s