重组Anti-S100A4抗体[EPR14639(2)] (ab197896)
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
- Suitable for: IHC-P, WB, ICC/IF, IP, Flow Cyt
- Knockout validated
- Reacts with: Mouse, Rat, Human
概述
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产品名称
Anti-S100A4抗体[EPR14639(2)]
参阅全部 S100A4 一抗 -
描述
兔单克隆抗体[EPR14639(2)] to S100A4 -
宿主
Rabbit -
特异性
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.
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Tested Applications & Species
Application Species Flow Cyt HumanICC/IF HumanIHC-P RatHumanIP HumanWB MouseHuman -
免疫原
Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: A549, A375, HeLa, NIH/3T3, Raw264.7 cell lysates. Human fetal spleen, colon and lung tissue lysates. Mouse spleen, bone marrow and heart tissues lysates. IHC-P: Human cervix carcinoma, lung carcinoma and gastric carcinoma tissues, rat spleen tissue. ICC/IF: HeLa cells. Flow Cyt: Jurkat cells.
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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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
性能
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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 long term. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR14639(2) -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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KO cell lines
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Recombinant Protein
应用
The Abpromise guarantee
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.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
应用 | Species |
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Flow Cyt |
Human
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ICC/IF |
Human
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IHC-P |
Rat
Human
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IP |
Human
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WB |
Mouse
Human
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应用 | Ab评论 | 说明 |
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IHC-P | (1) |
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).
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ICC/IF | (2) |
1/250.
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IP |
1/40.
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Flow Cyt |
1/250.
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说明 |
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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. |
靶标
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组织特异性
Ubiquitously expressed. -
序列相似性
Belongs to the S-100 family.
Contains 2 EF-hand domains. - Information by UniProt
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数据库链接
- Entrez Gene: 6275 Human
- Entrez Gene: 20198 Mouse
- Entrez Gene: 24615 Rat
- Omim: 114210 Human
- SwissProt: P26447 Human
- SwissProt: P07091 Mouse
- SwissProt: P05942 Rat
- Unigene: 654444 Human
see all -
别名
- 18A2 antibody
- 42A antibody
- calcium Placental protein antibody
see all
图片
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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 kDaLanes 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.
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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 kDaLanes 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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100A4 antibody [EPR14639(2)] (ab197896)
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).
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Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling S100A4 with ab197896 at 1/200 (10 μg/mL). Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% TritonX-100. ab150077, AlexaFluor®488 Goat anti-Rabbit at 1/1000 (2 μg/mL) was used as the secondary antibody. Cells were counterstained with ab195889, Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 (2.5 μg/mL). Nuclear counter stain was DAPI (blue).Confocal image showing positive staining in HeLa cell line.
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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 -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100A4 antibody [EPR14639(2)] (ab197896)
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.
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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 minutesBlocking/dilution buffer: 5% NFDM/TBST
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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.
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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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100A4 antibody [EPR14639(2)] (ab197896)
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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100A4 antibody [EPR14639(2)] (ab197896)
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.
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All lanes : Anti-S100A4 antibody [EPR14639(2)] (ab197896) at 1/1000 dilution
Lane 1 : A549 (Human lung carcinoma epithelial cell) whole cell lysates
Lane 2 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates
Lane 3 : Raw264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates
Lane 4 : Mouse spleen tissue lysates
Lane 5 : Mouse bone marrow tissue lysates
Lane 6 : Mouse heart tissue 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: 44 secondsBlocking and dilution buffer: 5% NFDM/TBST.
Some optimisation may be required for detection of the target protein due to low levels of endogenous expression in some samples.
实验方案
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (19)
ab197896 被引用在 19 文献中.
- Cai X et al. miR-124a enhances therapeutic effects of bone marrow stromal cells transplant on diabetic nephropathy-related epithelial-to-mesenchymal transition and fibrosis. J Cell Biochem 121:299-312 (2020). PubMed: 31190436
- Cai Y et al. Elimination of senescent cells by ß-galactosidase-targeted prodrug attenuates inflammation and restores physical function in aged mice. Cell Res N/A:N/A (2020). PubMed: 32341413
- Dhall S et al. A Flowable Placental Formulation Prevents Bleomycin-Induced Dermal Fibrosis in Aged Mice. Int J Mol Sci 21:N/A (2020). PubMed: 32545915
- Yin Y et al. Tongxinluo Attenuates Myocardiac Fibrosis after Acute Myocardial Infarction in Rats via Inhibition of Endothelial-to-Mesenchymal Transition. Biomed Res Int 2019:6595437 (2019). PubMed: 31317035
- Liu ZH et al. SIRT1 activation attenuates cardiac fibrosis by endothelial-to-mesenchymal transition. Biomed Pharmacother 118:109227 (2019). PubMed: 31351433