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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-S100A4 antibody [EPR14639(2)] - BSA and Azide free (ab220213)

Reviews (2) Submit a question References (2)
Western blot - Anti-S100A4 antibody [EPR14639(2)] - BSA and Azide free (ab220213)
  • Western blot - Anti-S100A4 antibody [EPR14639(2)] - BSA and Azide free (ab220213)
  • Western blot - Anti-S100A4 antibody [EPR14639(2)] - BSA and Azide free (ab220213)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100A4 antibody [EPR14639(2)] - BSA and Azide free (ab220213)
  • Immunoprecipitation - Anti-S100A4 antibody [EPR14639(2)] - BSA and Azide free (ab220213)
  • Immunoprecipitation - Anti-S100A4 antibody [EPR14639(2)] - BSA and Azide free (ab220213)
  • Flow Cytometry (Intracellular) - Anti-S100A4 antibody [EPR14639(2)] - BSA and Azide free (ab220213)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100A4 antibody [EPR14639(2)] - BSA and Azide free (ab220213)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100A4 antibody [EPR14639(2)] - BSA and Azide free (ab220213)
  • Anti-S100A4 antibody [EPR14639(2)] - BSA and Azide free (ab220213)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR14639(2)] to S100A4 - BSA and Azide free
  • Suitable for: Flow Cyt (Intra), IP, ICC/IF, WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Unconjugated

Overview

  • Product name

    Anti-S100A4 antibody [EPR14639(2)] - BSA and Azide free
    See all S100A4 primary antibodies

  • Description

    Rabbit monoclonal [EPR14639(2)] to S100A4 - BSA and Azide free

  • Host species

    Rabbit

  • 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.

  • Tested applications

    Suitable for: Flow Cyt (Intra), IP, ICC/IF, WB, IHC-Pmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HeLa, A549, and A375 cell lysates and human fetal spleen tissue lysates. IHC-P: Human cervix carcinoma, lung carcinoma and gastric carcinoma tissues. ICC/IF: Jurkat cells. Flow Cyt (intra): Jurkat cells.

  • General notes

    ab220213 is the carrier-free version of 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.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab220213 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
Flow Cyt (Intra)
Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
IP
Use at an assay dependent concentration.
ICC/IF
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Detects a band of approximately 12 kDa (predicted molecular weight: 12 kDa).
IHC-P (1)
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

We do not guarantee IHC-P for mouse and rat.
Notes
Flow Cyt (Intra)
Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
IP
Use at an assay dependent concentration.
ICC/IF
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Detects a band of approximately 12 kDa (predicted molecular weight: 12 kDa).
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.

We do not guarantee IHC-P for mouse and rat.

Target

  • Tissue specificity

    Ubiquitously expressed.

  • Sequence similarities

    Belongs to the S-100 family.
    Contains 2 EF-hand domains.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • 18A2 antibody
    • 42A antibody
    • calcium Placental protein antibody
    • Calvasculin antibody
    • CAPL antibody
    • Fibroblast specific protein 1 (FSP1) antibody
    • Fibroblast specific protein 1 antibody
    • Fibroblast specific protein antibody
    • FSP1 antibody
    • Leukemia multidrug resistance associated protein antibody
    • Malignant transformation suppression 1 (MTS1) antibody
    • Malignant transformation suppression 1 antibody
    • Metastasin antibody
    • MTS1 antibody
    • OTTHUMP00000015467 antibody
    • OTTHUMP00000015468 antibody
    • P9KA antibody
    • PEL98 antibody
    • Placental calcium-binding protein antibody
    • Protein Mts1 antibody
    • Protein S100 A4 antibody
    • Protein S100-A4 antibody
    • S100 calcium binding protein A4 (calcium protein, calvasculin, metastasin, murine placental homolog) antibody
    • S100 calcium binding protein A4 antibody
    • S100 calcium-binding protein A4 antibody
    • S100a4 antibody
    • S10A4_HUMAN antibody
    see all

Images

  • Western blot - Anti-S100A4 antibody [EPR14639(2)] - BSA and Azide free (ab220213)
    Western blot - Anti-S100A4 antibody [EPR14639(2)] - BSA and Azide free (ab220213)
    All lanes : Anti-S100A4 antibody [EPR14639(2)] (ab197896) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : S100A4 knockout HeLa cell lysate
    Lane 3 : Wild-type A549 cell lysate
    Lane 4 : S100A4 knockout A549 cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 12 kDa
    Observed band size: 11 kDa why is the actual band size different from the predicted?



    False colour image of Western blot: Anti-S100A4 antibody [EPR14639(2)] staining at 1/1000 dilution, shown in green; 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 ab265709 (knockout cell lysate ab257046) and S100A4 knockout A549 cell line ab261865 (knockout 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

  • Western blot - Anti-S100A4 antibody [EPR14639(2)] - BSA and Azide free (ab220213)
    Western blot - Anti-S100A4 antibody [EPR14639(2)] - BSA and Azide free (ab220213)
    All lanes : Anti-S100A4 antibody [EPR14639(2)] (ab197896) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : S100A4 knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 12 kDa
    Observed band size: 11 kDa why is the actual band size different from the predicted?



    This data was developed using the same antibody clone in a different buffer formulation (ab197896).

    Lanes 1 - 2: Merged signal (red and green). Green - ab197896 observed at 11 kDa. Red - 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 ab265709 (S100A4 knockout 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 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

  • Western blot - Anti-S100A4 antibody [EPR14639(2)] - BSA and Azide free (ab220213)
    Western blot - Anti-S100A4 antibody [EPR14639(2)] - BSA and Azide free (ab220213)
    All lanes : Anti-S100A4 antibody [EPR14639(2)] (ab197896) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : S100A4 knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 12 kDa
    Observed band size: 12 kDa



    This data was developed using the same antibody clone in a different buffer formulation (ab197896).

    Lanes 1 - 2: Merged signal (red and green). Green - ab197896 observed at 12 kDa. Red - 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 (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 (ab216773) and 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.

     

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100A4 antibody [EPR14639(2)] - BSA and Azide free (ab220213)
    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 ab197896 at 1/2000 dilution. 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab197896).

    Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunoprecipitation - Anti-S100A4 antibody [EPR14639(2)] - BSA and Azide free (ab220213)
    Immunoprecipitation - Anti-S100A4 antibody [EPR14639(2)] - BSA and Azide free (ab220213)

    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 (ab172730) instead of ab197896 in A549 whole cell lysate

    For western blotting, Panel A: ab197896, 1:1000; Panel B: 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 (ab197896).

  • Immunoprecipitation - Anti-S100A4 antibody [EPR14639(2)] - BSA and Azide free (ab220213)
    Immunoprecipitation - Anti-S100A4 antibody [EPR14639(2)] - BSA and Azide free (ab220213)

    ab197896 at 1/40 immunoprecipitating S100A4 in A549 whole cell lysate observed at 12 KDa.

    Lane 1 (+): ab197896 + A549 whole cell lysate.

    Lane 2 (-): Rabbit monoclonal IgG (ab172730) instead of ab197896 in A549 whole cell lysate

    For western blotting, 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 (ab197896).

  • Flow Cytometry (Intracellular) - Anti-S100A4 antibody [EPR14639(2)] - BSA and Azide free (ab220213)
    Flow Cytometry (Intracellular) - Anti-S100A4 antibody [EPR14639(2)] - BSA and Azide free (ab220213)

    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 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 (ab197896). 

     

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100A4 antibody [EPR14639(2)] - BSA and Azide free (ab220213)
    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 ab197896 at 1/2000 dilution. 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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)
    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 ab197896 at 1/2000 dilution. 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab197896).

    Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Anti-S100A4 antibody [EPR14639(2)] - BSA and Azide free (ab220213)
    Anti-S100A4 antibody [EPR14639(2)] - BSA and Azide free (ab220213)

References (2)

Publishing research using ab220213? Please let us know so that we can cite the reference in this datasheet.

ab220213 has been referenced in 2 publications.

  • Ageeli W  et al. Multimodality Characterization of Cancer-Associated Fibroblasts in Tumor Microenvironment and Its Correlation With Ultrasound Shear Wave-Measured Tissue Stiffness in Localized Prostate Cancer. Front Oncol 12:822476 (2022). PubMed: 35530322
  • Liu Y  et al. Extracellular ATP drives breast cancer cell migration and metastasis via S100A4 production by cancer cells and fibroblasts. Cancer Lett 430:1-10 (2018). PubMed: 29733962

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-S100A4 antibody [EPR14639(2)] - BSA and Azide free

 Excellent
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Lymph Node)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Tris Base, 1mM EDTA, pH 9.2
Permeabilization
No
Specification
Lymph Node
Blocking step
3% BSA, 5% horse serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 8% · Temperature: 20°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted May 31 2022

Mass Cytometry abreview for Anti-S100A4 antibody [EPR14639(2)] - BSA and Azide free

 Good
Abreviews
abreview image
Application
Mass Cytometry
Sample
Mouse Tissue sections (Lung mouse)
Specification
Lung mouse
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Dr. Tara Sutherland

Verified customer

Submitted Jul 20 2021

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