重组Anti-c-Kit抗体[YR145] (ab32363)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [YR145] to c-Kit
- Suitable for: IHC-P, WB
- Knockout validated
- Reacts with: Human, Common marmoset
Related conjugates and formulations
概述
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产品名称
Anti-c-Kit抗体[YR145]
参阅全部 c-Kit 一抗 -
描述
兔单克隆抗体[YR145] to c-Kit -
宿主
Rabbit -
特异性
This antibody is specific to c-Kit. It does not crossreact with other CSF1/PDGF receptor family member. -
经测试应用
适用于: IHC-P, WBmore details
不适用于: Flow Cyt or ICC/IF -
种属反应性
与反应: Human, Common marmoset -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: Human fetal kidney, fetal lung and seminoma tissue lysate. IHC-P: Human lung adenocarcinoma and seminoma tissues.
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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.
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. Avoid freeze / thaw cycle. -
解离常数(KD)
KD = 5.90 x 10 -11 M Learn more about KD -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
YR145 -
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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Positive Controls
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab32363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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IHC-P | (4) |
1/400. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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WB | (1) |
1/1000. Predicted molecular weight: 110 kDa.
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说明 |
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IHC-P
1/400. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
WB
1/1000. Predicted molecular weight: 110 kDa. |
靶标
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功能
Tyrosine-protein kinase that acts as cell-surface receptor for the cytokine KITLG/SCF and plays an essential role in the regulation of cell survival and proliferation, hematopoiesis, stem cell maintenance, gametogenesis, mast cell development, migration and function, and in melanogenesis. In response to KITLG/SCF binding, KIT can activate several signaling pathways. Phosphorylates PIK3R1, PLCG1, SH2B2/APS and CBL. Activates the AKT1 signaling pathway by phosphorylation of PIK3R1, the regulatory subunit of phosphatidylinositol 3-kinase. Activated KIT also transmits signals via GRB2 and activation of RAS, RAF1 and the MAP kinases MAPK1/ERK2 and/or MAPK3/ERK1. Promotes activation of STAT family members STAT1, STAT3, STAT5A and STAT5B. Activation of PLCG1 leads to the production of the cellular signaling molecules diacylglycerol and inositol 1,4,5-trisphosphate. KIT signaling is modulated by protein phosphatases, and by rapid internalization and degradation of the receptor. Activated KIT promotes phosphorylation of the protein phosphatases PTPN6/SHP-1 and PTPRU, and of the transcription factors STAT1, STAT3, STAT5A and STAT5B. Promotes phosphorylation of PIK3R1, CBL, CRK (isoform Crk-II), LYN, MAPK1/ERK2 and/or MAPK3/ERK1, PLCG1, SRC and SHC1. -
组织特异性
Isoform 1 and isoform 2 are detected in spermatogonia and Leydig cells. Isoform 3 is detected in round spermatids, elongating spermatids and spermatozoa (at protein level). Widely expressed. Detected in the hematopoietic system, the gastrointestinal system, in melanocytes and in germ cells. -
疾病相关
Piebald trait
Gastrointestinal stromal tumor
Testicular germ cell tumor
Leukemia, acute myelogenous -
序列相似性
Belongs to the protein kinase superfamily. Tyr protein kinase family. CSF-1/PDGF receptor subfamily.
Contains 5 Ig-like C2-type (immunoglobulin-like) domains.
Contains 1 protein kinase domain. -
翻译后修饰
Ubiquitinated by SOCS6. KIT is rapidly ubiquitinated after autophosphorylation induced by KITLG/SCF binding, leading to internalization and degradation.
Autophosphorylated on tyrosine residues. KITLG/SCF binding enhances autophosphorylation. Isoform 1 shows low levels of tyrosine phosphorylation in the absence of added KITLG/SCF (in vitro). Kinase activity is down-regulated by phosphorylation on serine residues by protein kinase C family members. Phosphorylation at Tyr-568 is required for interaction with PTPN11/SHP-2, CRK (isoform Crk-II) and members of the SRC tyrosine-protein kinase family. Phosphorylation at Tyr-570 is required for interaction with PTPN6/SHP-1. Phosphorylation at Tyr-703, Tyr-823 and Tyr-936 is important for interaction with GRB2. Phosphorylation at Tyr-721 is important for interaction with PIK3R1. Phosphorylation at Tyr-823 and Tyr-936 is important for interaction with GRB7. -
细胞定位
Cell membrane and Cytoplasm. Detected in the cytoplasm of spermatozoa, especially in the equatorial and subacrosomal region of the sperm head. - Information by UniProt
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数据库链接
- Entrez Gene: 3815 Human
- Omim: 164920 Human
- SwissProt: P10721 Human
- Unigene: 479754 Human
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别名
- C Kit antibody
- c-Kit antibody
- c-Kit Ligand antibody
see all
图片
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human seminoma tissue labelling c-Kit with unpurified ab32363.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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All lanes : Anti-c-Kit antibody [YR145] (ab32363) at 1/1000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : KIT knockout HAP1 whole cell lysate
Lane 3 : HUVEC whole cell lysate
Lysates/proteins at 40 µg per lane.
Predicted band size: 110 kDaLanes 1 - 3: Merged signal (red and green). Green - ab32363 observed at 109 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab32363 was shown to recognize 0 in wild-type HAP1 cells as signal was lost at the expected MW in KIT knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and KIT knockout samples were subjected to SDS-PAGE. Ab32363 and ab9484 (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) analysis of human seminoma tissue labelling c-Kit with purified ab32363 at 1/400. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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Anti-c-Kit antibody [YR145] (ab32363) at 1/5000 dilution (purified) + Human seminoma tissue lysate at 10 µg
Secondary
HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG at 1/50000 dilution
Predicted band size: 110 kDa
Observed band size: 120-145 kDa why is the actual band size different from the predicted?Blocking and dilution buffer: 5% NFDM/TBST.
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Anti-c-Kit antibody [YR145] (ab32363) at 1/1000 dilution (purified) + Human fetal lung tissue lysate at 10 µg
Secondary
HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG at 1/50000 dilution
Predicted band size: 110 kDa
Observed band size: 120-145 kDa why is the actual band size different from the predicted?Blocking and dilution buffer: 5% NFDM/TBST.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human glioma tissue labelling c-Kit with unpurified ab32363.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labelling c-Kit with unpurified ab32363.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human ovarian carcinoma tissue labelling c-Kit with unpurified ab32363.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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IHC-P image of c-Kit (unpurified ab32363) staining on adult marmoset testis. The sections were fixed in formaldehyde and underwent heat mediated antigen retrieval, using Dako antigen retrieval solution. The sections were then blocked in 5% milk for 30 mins at 25°C. The primary antibody was added at a 1/100 dilution, and then incubated for 18 hours at 4°C. Dapi was used to stain the nuclei (blue).
实验方案
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
数据表及文件
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SDS download
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Datasheet download
文献 (50)
ab32363 被引用在 50 文献中.
- Shima T et al. Glucose transporter‑1 inhibition overcomes imatinib resistance in gastrointestinal stromal tumor cells. Oncol Rep 47:N/A (2022). PubMed: 34738628
- Fang T et al. Evaluation of Immune Infiltration Based on Image Plus Helps Predict the Prognosis of Stage III Gastric Cancer Patients with Significantly Different Outcomes in Northeastern China. Dis Markers 2022:2893336 (2022). PubMed: 35371344
- Liu J et al. Exosomes from tamoxifen-resistant breast cancer cells transmit drug resistance partly by delivering miR-9-5p. Cancer Cell Int 21:55 (2021). PubMed: 33451320
- Yuan H et al. Premigratory neural crest stem cells generate enteric neurons populating the mouse colon and regulating peristalsis in tissue-engineered intestine. Stem Cells Transl Med 10:922-938 (2021). PubMed: 33481357
- Tang F et al. Patient-Derived Tumor Xenografts Plus Ex Vivo Models Enable Drug Validation for Tenosynovial Giant Cell Tumors. Ann Surg Oncol N/A:N/A (2021). PubMed: 33748895