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 APC Carrier Free HRP PE

产品名称

Anti-S100A4抗体[EPR2761(2)]
参阅全部 S100A4 一抗
描述

兔单克隆抗体[EPR2761(2)] to S100A4
宿主

Rabbit
特异性

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.
经测试应用

适用于: ICC/IF, Flow Cyt (Intra), WB, IP, IHC-Pmore details
种属反应性

与反应: Human
免疫原

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)
阳性对照
- Human tonsil, A549, A375, HeLa and Human small intestine lysates; Human tonsil tissue
常规说明
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.

形式

Liquid
存放说明

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.
存储溶液

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

Protein A purified
克隆

单克隆
克隆编号

EPR2761(2)
同种型

IgG
研究领域

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)

The Abpromise guarantee

Abpromise™承诺保证使用ab124805于以下的经测试应用

“应用说明”部分下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用	Ab评论	说明
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.

说明
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.

组织特异性

Ubiquitously expressed.
序列相似性

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
数据库链接
- Entrez Gene: 6275 Human
- Omim: 114210 Human
- SwissProt: P26447 Human
- Unigene: 654444 Human
别名
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- OTTHUMP00000015467 antibody
- OTTHUMP00000015468 antibody
- P9KA antibody
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- Placental calcium-binding protein antibody
- Protein Mts1 antibody
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- Protein S100-A4 antibody
- S100 calcium binding protein A4 (calcium protein, calvasculin, metastasin, murine placental homolog) antibody
- S100 calcium binding protein A4 antibody
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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

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发表研究结果有使用 ab124805？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab124805 被引用在 42 文献中.

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

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1-8 of 8 Abreviews or Q&A

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

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.

Luis Miguel Cardoso Dos Santos

Verified customer

提交于 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

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

提交于 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

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

提交于 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

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.

Hongwei Shao

Verified customer

提交于 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

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

提交于 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

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

提交于 Sep 26 2016

Application

Immunocytochemistry/ Immunofluorescence

Sample

Human Cell (A549)

Specification

A549

Fixative

Paraformaldehyde

Permeabilization

Yes - 0.25% Triton X-100/PBS

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

提交于 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

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

提交于 Jan 11 2013

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Key features and details

Related conjugates and formulations

概述

产品名称

描述

宿主

特异性

经测试应用

种属反应性

免疫原

阳性对照

常规说明

性能

形式

存放说明

存储溶液

纯度

克隆

克隆编号

同种型

研究领域

相关产品

Alternative Versions

Compatible Secondaries

Isotype control

KO cell lines

KO cell lysates

Positive Controls

应用

The Abpromise guarantee

靶标

组织特异性

序列相似性

数据库链接

别名

图片

实验方案

数据表及文件

SDS download

Datasheet download

文献 (42)

客户评价及客户问答

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)]