重组Anti-S100A4抗体[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
Related conjugates and formulations
概述
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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.
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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
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阳性对照
- Human tonsil, A549, A375, HeLa and Human small intestine lysates; Human tonsil tissue
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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.
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.
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. 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...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR2761(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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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
应用
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 |
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ICC/IF | (2) |
1/100 - 1/250.
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Flow Cyt (Intra) |
1/80 - 1/800.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB | (1) |
1/1000 - 1/10000. Detects a band of approximately 12 kDa (predicted molecular weight: 12 kDa).
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IP |
1/10 - 1/100.
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IHC-P | (4) |
1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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说明 |
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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. |
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
- Omim: 114210 Human
- SwissProt: P26447 Human
- Unigene: 654444 Human
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别名
- 18A2 antibody
- 42A antibody
- calcium Placental protein antibody
see all
图片
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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.
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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 kDaLanes 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.
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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.
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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)
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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)
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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.
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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 -
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 -
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
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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
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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.
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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. -
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)
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (42)
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