Name: Anti-S100A4 antibody [EPR14639(2)] - BSA and Azide free
SKU: ab220213
Rating: 5 (2 reviews)

Rabbit Recombinant Monoclonal S100A4 antibody. Carrier free. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human, Mouse, Rat samples. Cited in 2 publications.

Images

Western blot - Anti-S100A4 antibody [EPR14639(2)] - BSA and Azide free (AB220213), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: PBS
Form: Liquid
Clonality: Monoclonal

Immunogen

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

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

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

Tested
Tested

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

Predicted
Predicted

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Predicted
Predicted

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info -	Notes -

Expected
Expected

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

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Predicted
Predicted

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	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.

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.

Associated Products

Select an associated product type

9 products for Alternative Product

1 product for Alternative Version

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

Recommended products

Rabbit Recombinant Monoclonal S100A4 antibody. Carrier free. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human, Mouse, Rat samples. Cited in 2 publications.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: PBS
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Carrier free: Yes
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 conditions: +4°C
Appropriate long-term storage conditions: +4°C
Storage information: Do Not Freeze

Notes

ab220213 is the carrier-free version of Anti-S100A4 antibody [EPR14639(2)] ab197896.

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.

Supplementary info

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

Activity summary

S100A4 also known as metastasin or MTS1 is a protein with a molecular weight of approximately 12 kDa. It functions by binding calcium which induces conformational changes aiding in its interaction with target proteins. S100A4 is part of the S100 family of proteins characterized by two EF-hand calcium-binding motifs. Expression of S100A4 is observed in various cell types including fibroblasts endothelial cells and several tumor cells where it contributes to processes like cell motility invasion and angiogenesis.

Biological function summary

The role of S100A4 extends beyond mere calcium binding; it associates with the cytoskeleton promoting cell structures related to movement and metastasis. It does not act alone but often partners with proteins like myosin IIA to regulate cytoskeletal dynamics. Additionally S100A4 modulates extracellular matrix components and interacts with signaling pathways that control cancer progression making it significant in both normal physiological contexts and pathological states.

Pathways

Functional activities of S100A4 integrate into important pathways involved in cellular movement and metastasis. S100A4 plays roles in the regulation of the Wnt/β-catenin pathway which is pivotal for cell-cell interaction and fate determination. It also interacts with matrix metalloproteinases like MMPs thereby managing processes of tissue invasion and remodeling essential in cancer metastasis. Other pathways influencing cell adhesion and signal transduction may indirectly involve S100A4 through related proteins.

Associated diseases and disorders

Elevated S100A4 levels have implications in cancer particularly in enhancing metastasis in breast and colorectal cancers. The protein promotes tumor developments by inducing changes in the tumor microenvironment making it a target of interest in oncology. S100A4 also interacts with proteins like p53 and β-catenin which are critical in the pathology of tumors. Moreover S100A4 contributes to fibrotic diseases accentuating tissue remodeling activities by modulating fibroblast behavior and extracellular matrix deposition.

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.

10 product images

Western blot - Anti-S100A4 antibody [EPR14639(2)] - BSA and Azide free (ab220213)
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, Anti-S100A4 antibody [EPR14639(2)] 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)] (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)] - BSA and Azide free (ab220213)
This data was developed using the same antibody clone in a different buffer formulation (Anti-S100A4 antibody [EPR14639(2)] ab197896).
Lanes 1 - 2: Merged signal (red and green). Green - Anti-S100A4 antibody [EPR14639(2)] 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.
Anti-S100A4 antibody [EPR14639(2)] 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 Anti-S100A4 antibody [EPR14639(2)] 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)] (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
Immunoprecipitation - Anti-S100A4 antibody [EPR14639(2)] - BSA and Azide free (ab220213)
Anti-S100A4 antibody [EPR14639(2)] ab197896 at 1/40 immunoprecipitating S100A4 in A549 whole cell lysate observed at 12 KDa.
Lane 1 (+): Anti-S100A4 antibody [EPR14639(2)] ab197896 + A549 whole cell lysate.
Lane 2 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-S100A4 antibody [EPR14639(2)] ab197896 in A549 whole cell lysate
For western blotting, Anti-S100A4 antibody [EPR14639(2)] ab197896 at 1/1000 and anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG (1/1500).
Blocking/Diluting buffer and concentration: 5% NFDM/TBST.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-S100A4 antibody [EPR14639(2)] ab197896).
All lanes: Immunoprecipitation - Anti-S100A4 antibody [EPR14639(2)] - BSA and Azide free (ab220213)
Predicted band size: 12 kDa
Western blot - Anti-S100A4 antibody [EPR14639(2)] - BSA and Azide free (ab220213)
This data was developed using the same antibody clone in a different buffer formulation (Anti-S100A4 antibody [EPR14639(2)] ab197896).
Lanes 1 - 2: Merged signal (red and green). Green - Anti-S100A4 antibody [EPR14639(2)] ab197896 observed at 12 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 37 kDa.
Anti-S100A4 antibody [EPR14639(2)] 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. Anti-S100A4 antibody [EPR14639(2)] 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)] (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
Immunoprecipitation - Anti-S100A4 antibody [EPR14639(2)] - BSA and Azide free (ab220213)
Anti-S100A4 antibody [EPR14639(2)] 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 (+): Anti-S100A4 antibody [EPR14639(2)] ab197896 + A549 whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-S100A4 antibody [EPR14639(2)] ab197896 in A549 whole cell lysate
For western blotting, Panel A: Anti-S100A4 antibody [EPR14639(2)] 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-S100A4 antibody [EPR14639(2)] ab197896).
All lanes: Immunoprecipitation - Anti-S100A4 antibody [EPR14639(2)] (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)] - BSA and Azide free (ab220213)
Immunohistochemical analysis of paraffin-embedded Human gastric carcinoma tissue labeling S100A4 using Anti-S100A4 antibody [EPR14639(2)] 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 Anti-S100A4 antibody [EPR14639(2)] ab197896 and secondary antibody only.
Note: Cytoplasm and nuclear staining on human gastric carcinoma tissue was observed.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-S100A4 antibody [EPR14639(2)] ab197896).
Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cytometry (Intracellular) - Anti-S100A4 antibody [EPR14639(2)] - BSA and Azide free (ab220213)
Intracellular Flow Cytometry analysis of Jurkat cells labelling S100A4 with Anti-S100A4 antibody [EPR14639(2)] ab197896 at 1/250 (red). Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. An Alexa Fluor^® 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-S100A4 antibody [EPR14639(2)] ab197896).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100A4 antibody [EPR14639(2)] - BSA and Azide free (ab220213)
Immunohistochemical analysis of paraffin-embedded Human cervix carcinoma tissue labeling S100A4 using Anti-S100A4 antibody [EPR14639(2)] 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 Anti-S100A4 antibody [EPR14639(2)] ab197896 and secondary antibody only.
Note: Nuclear and cytoplasm staining on cervix carcinoma tissue was observed.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-S100A4 antibody [EPR14639(2)] ab197896).
Heat mediated antigen retrieval was performed 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)] - BSA and Azide free (ab220213)
Immunohistochemical analysis of paraffin-embedded Human lung carcinoma tissue labeling S100A4 using Anti-S100A4 antibody [EPR14639(2)] 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 Anti-S100A4 antibody [EPR14639(2)] ab197896 and secondary antibody only.
Note: Nuclear and weakly staining on lung carcinoma tissue was observed.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-S100A4 antibody [EPR14639(2)] ab197896).
Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-S100A4 antibody [EPR14639(2)] - BSA and Azide free (ab220213)
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-S100A4 antibody [EPR14639(2)] ab197896).
Blocking and diluting buffer and concentration: 5% NFDM /TBST.
Compared with Anti-S100A4 antibody [EPR14639(2)] 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 Anti-S100A4 antibody [EPR14639(2)] ab197896 in western blot.
All lanes: Western blot - Anti-S100A4 antibody [EPR14639(2)] (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