Name: Anti-S100A4 antibody [EPR2761(2)]
Brand: RabMAb
SKU: ab124805
Rating: 4 (8 reviews)

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Western blot - Anti-S100A4 antibody [EPR2761(2)] (ab124805)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR2761(2)] to S100A4
Suitable for: ICC/IF, Flow Cyt (Intra), WB, IP, IHC-P
Knockout validated
Reacts with: Human

Alexa Fluor® 488 Alexa Fluor® 647 Carrier Free

Product name

Anti-S100A4 antibody [EPR2761(2)]
See all S100A4 primary antibodies
Description

Rabbit monoclonal [EPR2761(2)] to S100A4
Host species

Rabbit
Specificity

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.
Tested applications

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

Reacts with: Human
Immunogen

Synthetic peptide within Human S100A4 aa 50 to the C-terminus (C terminal). The exact sequence is proprietary.
Database link: P26447
(Peptide available as ab232852)
Positive control
- Human tonsil, A549, A375, HeLa and Human small intestine lysates; Human tonsil tissue
General notes
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 production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR2761(2)
Isotype

IgG
Research areas

Alternative Versions
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
KO cell lines
- Human S100A4 knockout A549 cell line (ab261865)
- Human S100A4 knockout HeLa cell line (ab265709)
KO cell lysates
- Human S100A4 knockout HeLa cell lysate (ab257046)
- Human S100A4 knockout A549 cell lysate (ab261674)
Positive Controls
- A549 whole cell lysate (ab7910)
Recombinant Protein
- Recombinant Human S100A4 protein (ab85489)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab124805 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	(2)	1/100 - 1/250.
Flow Cyt (Intra)		1/80 - 1/800. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
WB	(1)	1/1000 - 1/10000. Detects a band of approximately 12 kDa (predicted molecular weight: 12 kDa).
IP		1/10 - 1/100.
IHC-P	(4)	1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Notes
ICC/IF 1/100 - 1/250.
Flow Cyt (Intra) 1/80 - 1/800. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
WB 1/1000 - 1/10000. Detects a band of approximately 12 kDa (predicted molecular weight: 12 kDa).
IP 1/10 - 1/100.
IHC-P 1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Tissue specificity

Ubiquitously expressed.
Sequence similarities

Belongs to the S-100 family.
Contains 2 EF-hand domains.
Target information above from: UniProt accession P26447 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 6275 Human
- Omim: 114210 Human
- SwissProt: P26447 Human
- Unigene: 654444 Human
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

Western blot - Anti-S100A4 antibody [EPR2761(2)] (ab124805)

All lanes : Anti-S100A4 antibody [EPR2761(2)] (ab124805) 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?

Lanes 1 - 2: Merged signal (red and green). Green - ab124805 observed at 11 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

ab124805 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 ab124805 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 [EPR2761(2)] (ab124805)

All lanes : Anti-S100A4 antibody [EPR2761(2)] (ab124805) at 1/1000 dilution

Lane 1 : Wild-type A549 (Human lung carcinoma cell line) whole cell lysate
Lane 2 : S100A4 knockout A549 (Human lung carcinoma cell line) whole cell lysate
Lane 3 : A-375 (Human malignant melanoma cell line) whole cell lysate
Lane 4 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

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

Lanes 1 - 4: Merged signal (red and green). Green - ab124805 observed at 12 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

ab124805 was shown to react with S100A4 in wild-type A549 cells in Western blot with loss of signal observed in S100A4 knockout cell line ab261865 (knockout cell lysate ab261674). Wild-type A549 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 ab124805 and ab8245 (Mouse anti-GAPDH antibody [6C5]) 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.
Immunocytochemistry/ Immunofluorescence - Anti-S100A4 antibody [EPR2761(2)] (ab124805)This image is courtesy of an anonymous Abreview

Unpurified ab124805 staining S100A4 in the A549 cell line from Human lungs by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and permeabilized with Triton X-100 0.25% in PBS. Samples were incubated with primary antibody (1/100) for 45 minutes at 25°C. A TRITC-conjugated Goat anti-rabbit polyclonal was used as the secondary antibody.
See Abreview
Flow Cytometry (Intracellular) - Anti-S100A4 antibody [EPR2761(2)] (ab124805)

ab124805 staining S100A4 in HeLa (human cervix adenocarcinoma) by intracellular flow cytometry. Cells were fixed with 80% methanoland permeabilised with 0.1% Triton X-100 (in PBS). The sample was incubated with the primary antibody at a dilution of 1/800. A goat anti rabbit IgG (Alexa Fluor^® 488) at a dilution of 1/2000 was used as the secondary antibody.

Isoytype control: ab172730 rabbit monoclonal IgG (Black)

Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)
Flow Cytometry (Intracellular) - Anti-S100A4 antibody [EPR2761(2)] (ab124805)

ab124805 staining S100A4 in HeLa (human cervix adenocarcinoma) by intracellular flow cytometry. Cells were fixed with 80% methanoland permeabilised with 0.1% Triton X-100 (in PBS). The sample was incubated with the primary antibody at a dilution of 1/80. A goat anti rabbit IgG (Alexa Fluor^® 488) at a dilution of 1/2000 was used as the secondary antibody.

Isoytype control: ab172730 rabbit monoclonal IgG (Black)

Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100A4 antibody [EPR2761(2)] (ab124805)

ab124805 staining S100A4 in human tonsil tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/500. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

Negative control 1: PBS in place of primary antibody.
Western blot - Anti-S100A4 antibody [EPR2761(2)] (ab124805)

All lanes : Anti-S100A4 antibody [EPR2761(2)] (ab124805) at 1/1000 dilution (unpurified)

Lane 1 : Human tonsil lysate
Lane 2 : A549 cell lysate
Lane 3 : A375 cell lysate
Lane 4 : Human small intestine lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 12 kDa
Observed band size: 12 kDa
Immunoprecipitation - Anti-S100A4 antibody [EPR2761(2)] (ab124805)

ab124805 immunoprecipitating S100A4. 10µg of cell lysate was incubated with primary antibody at a dilution of 1/30 and Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at a dilution of 1/1500.

Lane 1: Human tonsil whole cell lysate (10ug)
Lane 2: Human tonsil whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab124805 in human tonsil whole cell lysate
Western blot - Anti-S100A4 antibody [EPR2761(2)] (ab124805)

Anti-S100A4 antibody [EPR2761(2)] (ab124805) + A549 (human lung carcinoma) whole cell lysate at 20 µg

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

Predicted band size: 12 kDa
Additional bands at: 12 kDa. We are unsure as to the identity of these extra bands.

Blocking buffer: 5% NFDM /TBST

Diluting Buffer: 5% NFDM /TBST
Western blot - Anti-S100A4 antibody [EPR2761(2)] (ab124805)

All lanes : Anti-S100A4 antibody [EPR2761(2)] (ab124805) at 1/2500 dilution

Lane 1 : Human tonsil tissue lysate
Lane 2 : A375 (human malignant melanoma) whole cell 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
Additional bands at: 12 kDa. We are unsure as to the identity of these extra bands.

Blocking Buffer: 5% NFDM /TBST

Diluting Buffer: 5% NFDM /TBST
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100A4 antibody [EPR2761(2)] (ab124805)

Unpurified ab124805, at 1/250 dilution, staining S100A4 in paraffin-embedded Human tonsil tissue by Immunohistochemistry.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100A4 antibody [EPR2761(2)] (ab124805)This image is courtesy of an anonymous Abreview

Immunohistochemical analysis of Human lung tissue, staining S100A4 with unpurified ab124805.

Tissue was fixed with HOPE and blocked with blocking solution for 5 minutes at 25°C. Samples were incubated with primary antibody (1/1000 in diluent) for 1 hour at 25°C. An undiluted HRP-conjugated goat anti-rabbit polyclonal IgG was used as the secondary antibody.
See Abreview
Immunocytochemistry/ Immunofluorescence - Anti-S100A4 antibody [EPR2761(2)] (ab124805)

ab124805 staining S100A4 in Jurkat (human acute T cell leukemia) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/250. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at 1/500. ab7291 and ab150120 were used as counterstains for primary antibody ab124805 and secondary antibody ab150077 respectively and DAPI was used as a nuclear counterstain.

Negative control 1: Rabbit primary antibody and anti-mouse secondary antibody (ab150120)
Negative control 2: Mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab150077)
Anti-S100A4 antibody [EPR2761(2)] (ab124805)

Flow cytometry protocols
Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

Publishing research using ab124805? Please let us know so that we can cite the reference in this datasheet.

ab124805 has been referenced in 42 publications.

Cao X et al. MSC-derived exosomal lncRNA SNHG7 suppresses endothelial-mesenchymal transition and tube formation in diabetic retinopathy via miR-34a-5p/XBP1 axis. Life Sci 272:119232 (2021). PubMed: 33600866
Singh M et al. 22(R)-hydroxycholesterol for dopaminergic neuronal specification of MSCs and amelioration of Parkinsonian symptoms in rats. Cell Death Discov 7:13 (2021). PubMed: 33454721
Lyu T et al. Exosomes from BM-MSCs promote acute myeloid leukemia cell proliferation, invasion and chemoresistance via upregulation of S100A4. Exp Hematol Oncol 10:24 (2021). PubMed: 33789743
Zhang HW et al. Cancer-associated fibroblast-derived exosomal microRNA-24-3p enhances colon cancer cell resistance to MTX by down-regulating CDX2/HEPH axis. J Cell Mol Med 25:3699-3713 (2021). PubMed: 33621425
Mussa BM et al. Differentiated PDGFRa-Positive Cells: A Novel In-Vitro Model for Functional Studies of Neuronal Nitric Oxide Synthase. Int J Mol Sci 22:N/A (2021). PubMed: 33805311

View all Publications for this product

Customer reviews and Q&As

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1-8 of 8 Abreviews

Application

Western blot

Sample

Mouse Cell lysate - whole cell (smooth muscle cells)

Gel Running Conditions

Reduced Denaturing (12% gel)

Loading amount

10 µg

Specification

smooth muscle cells

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Luis Miguel Cardoso Dos Santos

Verified customer

Submitted Feb 24 2022

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Rhesus monkey Tissue sections (Tonsil)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Citrate buffer

Permeabilization

Specification

Tonsil

Blocking step

Leica IHC Super blocking as blocking agent for 20 minute(s) · Concentration: 100% · Temperature: 22°C

Fixative

Paraformaldehyde

Abcam user community

Verified customer

Submitted Jun 01 2018

Application

Immunocytochemistry/ Immunofluorescence

Sample

Rhesus monkey Cell (Dermal Fibroblast)

Permeabilization

Yes - 0.02% Triton X-100 with 0.1mg/mL BSA in PBS, 7 min

Specification

Dermal Fibroblast

Blocking step

BSA as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 20°C

Fixative

Paraformaldehyde

Abcam user community

Verified customer

Submitted Feb 23 2018

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Mouse Tissue sections (skin)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Sodium Citrate pH6.0

Permeabilization

Yes - 0.25%Triton X100 in PBS

Specification

skin

Blocking step

Dako X0909 as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: RT°C

Fixative

Formaldehyde

Hongwei Shao

Verified customer

Submitted Sep 29 2017

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Human Tissue sections (Skin)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: pH9

Permeabilization

Specification

Skin

Blocking step

Serum as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 21°C

Fixative

Paraformaldehyde

Abcam user community

Verified customer

Submitted Mar 06 2017

Application

Immunohistochemistry (Frozen sections)

Sample

Human Tissue sections (Skin)

Permeabilization

Specification

Skin

Blocking step

Casein as blocking agent for 10 minute(s) · Concentration: 0.25% · Temperature: 23°C

Fixative

Acetone

Abcam user community

Verified customer

Submitted Sep 26 2016

Application

Immunocytochemistry/ Immunofluorescence

Sample

Human Cell (A549)

Specification

A549

Fixative

Paraformaldehyde

Permeabilization

Yes - 0.25% Triton X-100/PBS

Abcam user community

Verified customer

Submitted Apr 05 2013

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Human Tissue sections (Lung tissue)

Specification

Lung tissue

Fixative

HOPE

Antigen retrieval step

None

Permeabilization

Blocking step

Zytomed Systems HRP Polymer Block Solution as blocking agent for 5 minute(s) · Concentration: 100% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jan 11 2013

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Specificity

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

KO cell lines

KO cell lysates

Positive Controls

Recombinant Protein

Applications

The Abpromise guarantee

Target

Tissue specificity

Sequence similarities

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

References (42)

Customer reviews and Q&As

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Western blot abreview for Anti-S100A4 antibody [EPR2761(2)]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-S100A4 antibody [EPR2761(2)]

Immunocytochemistry/ Immunofluorescence abreview for Anti-S100A4 antibody [EPR2761(2)]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-S100A4 antibody [EPR2761(2)]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-S100A4 antibody [EPR2761(2)]

Immunohistochemistry (Frozen sections) abreview for Anti-S100A4 antibody [EPR2761(2)]

Immunocytochemistry/ Immunofluorescence abreview for Anti-S100A4 antibody [EPR2761(2)]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-S100A4 antibody [EPR2761(2)]