Overview

  • Product name

    Anti-S100A4 antibody [EPR14639(2)]
    See all S100A4 primary antibodies
  • Description

    Rabbit monoclonal [EPR14639(2)] to S100A4
  • 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: IHC-P, WB, ICC/IF, IP, Flow Cytmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant full length protein corresponding to Human S100A4 aa 1 to the C-terminus.
    Database link: P26447

  • Positive control

    • WB: A549, A375, HeLa, Human fetal spleen, colon and lung lysates. IHC-P: Human cervix carcinoma, lung carcinoma and gastric carcinoma tissues, rat spleen tissue. ICC/IF: Jurkat cells. Flow Cyt: Jurkat cells.
  • General 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 rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab197896 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
IHC-P 1/2000. 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.

WB 1/1000. Detects a band of approximately 12 kDa (predicted molecular weight: 12 kDa).
ICC/IF 1/250.
IP 1/40.
Flow Cyt 1/250.

Target

  • Tissue specificity

    Ubiquitously expressed.
  • Sequence similarities

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

  • 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

  • All lanes : Anti-S100A4 antibody [EPR14639(2)] (ab197896) at 1/1000 dilution

    Lane 1 : Wild-type A549 whole cell lysate
    Lane 2 : S100A4 knockout A549 whole cell lysate
    Lane 3 : A375 whole cell lysate
    Lane 4 : HeLa whole cell lysate

    Lysates/proteins at 20 µg per lane.

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



    Lanes 1 - 4: Merged signal (red and green). Green - ab197896 observed at 12 kDa. Red - 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 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 ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemical analysis of paraffin-embedded Rat spleen  tissue labeling S100A4 with ab197896 at 1/2000 dilution (0.376 μg/ml)  followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on lymphocytes of rat spleen.The section was incubated with ab197896 for 30 mins at room temperature. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

  • All lanes : Anti-S100A4 antibody [EPR14639(2)] (ab197896) at 1/1000 dilution

    Lanes 1 & 5 : A549 whole cell lysate
    Lane 2 : Human liver lysates
    Lane 3 : Human lung lysates
    Lane 4 : Human colon lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

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


    Exposure time: 26 seconds
  • 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.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • All lanes : Anti-S100A4 antibody [EPR14639(2)] (ab197896) at 1/1000 dilution

    Lane 1 : A375 (human malignant melanoma) whole cell lysate
    Lane 2 : Human fetal spleen lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 12 kDa


    Exposure time: 3 minutes


    Blocking/dilution buffer: 5% NFDM/TBST

  • 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.

  • 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.

  • 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.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Anti-S100A4 antibody [EPR14639(2)] (ab197896) at 1/5000 dilution + A549 (Human lung carcinoma) whole cell lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 12 kDa


    Exposure time: 3 minutes


    Blocking/dilution buffer: 5% NFDM/TBST

  • 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.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • 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.

References

This product has been referenced in:

  • Wang M  et al. Novel inhibitors of the cellular renin-angiotensin system components, poricoic acids, target Smad3 phosphorylation and Wnt/ß-catenin pathway against renal fibrosis. Br J Pharmacol 175:2689-2708 (2018). Read more (PubMed: 29679507) »
  • Han WQ  et al. Membrane rafts-redox signalling pathway contributes to renal fibrosis via modulation of the renal tubular epithelial-mesenchymal transition. J Physiol 596:3603-3616 (2018). Read more (PubMed: 29863758) »
See all 3 Publications for this product

Customer reviews and Q&As

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Skin)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate
Permeabilization
No
Specification
Skin
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
Fixative
Paraformaldehyde

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Submitted Mar 03 2017

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