重组Anti-CD10抗体[EPR22865-73] (ab255609)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22865-73] to CD10
- Suitable for: WB, IHC-P, ICC/IF, IP, mIHC
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-CD10抗体[EPR22865-73]
参阅全部 CD10 一抗 -
描述
兔单克隆抗体[EPR22865-73] to CD10 -
宿主
Rabbit -
特异性
IHC application is recommended for human only.
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经测试应用
适用于: WB, IHC-P, ICC/IF, IP, mIHCmore details
不适用于: Flow Cyt -
种属反应性
与反应: Mouse, Rat, Human -
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 placenta tisssue lysate; rat kidney and liver lysates; Raji, Ramos and HAP1 whole cell lysates; rat kidney and liver lysates; mouse kidney lysate. IHC-P: Human kidney, breast, liver and tonsil tissue, human diffuse large B-cell lymphoma ICC/IF: Raji and Ramos cells. IP: Raji whole cell lysate. mIHC: Human breast and endometrium 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.
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: 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
EPR22865-73 -
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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Immunohistochemistry reagents
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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab255609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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WB |
1/1000. Detects a band of approximately 100 kDa (predicted molecular weight: 85 kDa).
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IHC-P |
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IHC application is recommended for human only. |
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ICC/IF |
1/50.
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IP |
1/30.
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mIHC |
1/1000.
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说明 |
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WB
1/1000. Detects a band of approximately 100 kDa (predicted molecular weight: 85 kDa). |
IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. IHC application is recommended for human only. |
ICC/IF
1/50. |
IP
1/30. |
mIHC
1/1000. |
靶标
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功能
Thermolysin-like specificity, but is almost confined on acting on polypeptides of up to 30 amino acids. Biologically important in the destruction of opioid peptides such as Met- and Leu-enkephalins by cleavage of a Gly-Phe bond. Able to cleave angiotensin-1, angiotensin-2 and angiotensin 1-9. Involved in the degradation of atrial natriuretic factor (ANF). Displays UV-inducible elastase activity toward skin preelastic and elastic fibers. -
序列相似性
Belongs to the peptidase M13 family. -
细胞定位
Cell membrane. - Information by UniProt
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数据库链接
- Entrez Gene: 4311 Human
- Entrez Gene: 17380 Mouse
- Entrez Gene: 24590 Rat
- Omim: 120520 Human
- SwissProt: P08473 Human
- SwissProt: Q61391 Mouse
- SwissProt: P07861 Rat
- Unigene: 307734 Human
see all -
别名
- Atriopeptidase antibody
- CALLA antibody
- CD10 antibody
see all
图片
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Fluorescence multiplex immunohistochemical analysis of the human endometrium (Formalin/PFA-fixed paraffin-embedded sections).
Panel A: merged staining of anti-E Cadherin (ab256580, red; Opal™690), anti-SLC34A2 (ab238793, green; Opal™520) and anti-CD10 (ab255609, cyan; Opal™570) on human endometrium. Panel B: anti-CD10 stained on stromal cells. Panel C: anti-E Cadherin stained on glandular cells. Panel D: anti-SLC34A2 stained on apical membrane of glandular cells. Opal Polymer HRP Ms + Rb was used as a secondary antibody.
The section was incubated in three rounds of staining: in the order of ab256580 at 1/3000 dilution (0.324 μg/ml) for 30mins, ab238793 at 1/1000 dilution (2.26 μg/ml) for 10mins and ab255609 at 1/1000 dilution (0.615 μg/ml) for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. DAPI (blue) was used as a nuclear counter stain.
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Fluorescence multiplex immunohistochemical analysis of the human breast (Formalin/PFA-fixed paraffin-embedded sections).
Panel A: merged staining of anti-B7H4 (ab252438, red; Opal™690), anti-CD10 (ab255609, gray; Opal™520) and anti-FABP4 (ab92501, cyan; Opal™570) on human breast. Panel B: anti-B7H4 stained on glandular lumens. Panel C: anti-CD10 stained on myoepithelial cells. Panel D: anti-FABP4 stained on adipocytes. Opal Polymer HRP Ms + Rb was used as a secondary antibody.
The section was incubated in three rounds of staining: in the order of ab252438 at 1/100 dilution (4.69 μg/ml), ab255609 at 1/1000 dilution (0.615 μg/ml) and ab92501 at 1/10000 dilution (0.047 μg/ml) for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. DAPI (blue) was used as a nuclear counter stain.
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All lanes : Anti-CD10 antibody [EPR22865-73] (ab255609) at 1/1000 dilution
Lane 1 : Human placenta tissue lysate
Lane 2 : Raji (human Burkitt's lymphoma cell line) whole cell lysate
Lane 3 : Ramos (human Burkitt's lymphoma cell line) whole cell lysate
Lane 4 : HT-29 (human colorectal adenocarcinoma cell line) whole cell lysate
Lane 5 : Wild-type HAP1 whole cell lysate
Lane 6 : CD10 knockout HAP1 whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
Lane 1 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Lanes 2-4 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 85 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?ab255609 was shown to specifically react with CD10 in wild-type HAP1 cells as signal was lost in CD10 knockout cells. Wild-type and CD10 knockout samples were subjected to SDS-PAGE. ab255609 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument using the ECL technique.
Lanes 5-6 in this blot were developed using a higher sensitivity ECL substrate.
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure times.
Lane 1: 10 seconds; Lanes 2-4: 15 seconds; Lanes 5-6: 70 seconds.
The molecular weight observed is consistent with what has been described in the literature (PMID:15286660).
Negative control: HT-29 (PMID:19828468).
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CD10 was immunoprecipitated from 0.35 mg of Raji (human Burkitt's lymphoma cell line) whole cell lysate with ab255609 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab255609 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used as secondary antibody at 1/5000 dilution.
Lane 1: Raji whole cell lysate 10 μg (Input).
Lane 2: ab255609 IP in Raji whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab255609 in Raji whole cell lysate.Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 seconds. -
Immunofluorescent analysis of 100% methanol-fixed Ramos (human Burkitt's lymphoma cell line) cells labeling CD10 with ab255609 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic and membranous staining in Ramos cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
100% methanol was preferred as fixative.
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Immunofluorescent analysis of 100% methanol-fixed Raji (human Burkitt's lymphoma cell line) cells labeling CD10 with ab255609 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic and membranous staining in Raji cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
100% methanol was preferred as fixative.
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Anti-CD10 antibody [EPR22865-73] (ab255609) at 1/1000 dilution + Mouse kidney tissue lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 85 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking and dilution buffer: 5% NFDM/TBST.
The molecular weight observed is consistent with what has been described in the literature (PMID:15286660).
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Immunohistochemical analysis of paraffin-embedded Human diffuse large B-cell lymphoma labelling CD10 with ab255609 at 1/1000 dilution, followed by a Goat Anti-Rabbit IgG H&L (HRP polymer) ready to use (ab214880). Positive staining on human diffuse large B-cell lymphoma is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a Goat Anti-Rabbit IgG H&L (HRP polymer) ready to use.
Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
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All lanes : Anti-CD10 antibody [EPR22865-73] (ab255609) at 1/1000 dilution
Lane 1 : Rat kidney tissue lysate
Lane 2 : Rat liver tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 85 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?
Exposure time: 15 secondsBlocking and dilution buffer: 5% NFDM/TBST.
The molecular weight observed is consistent with what has been described in the literature (PMID:15286660).
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Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling CD10 with ab255609 at 1/1000 dilution, followed by a Goat Anti-Rabbit IgG H&L (HRP) ready to use. Membranous staining on germinal center of human tonsil (PMID:10843287) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a Goat Anti-Rabbit IgG H&L (HRP) ready to use.
Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
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Immunohistochemical analysis of paraffin-embedded human liver tissue labeling CD10 with ab255609 at 1/1000 dilution, followed by a Goat Anti-Rabbit IgG H&L (HRP) ready to use. Membranous staining on intrahepatic canaliculi of human liver (PMID:10705818) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a Goat Anti-Rabbit IgG H&L (HRP) ready to use.
Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
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Immunohistochemical analysis of paraffin-embedded human breast tissue labeling CD10 with ab255609 at 1/1000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP) ready to use. Membranous staining on myoepithelial cells of human breast (PMID:10705818, 17143263) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a Goat Anti-Rabbit IgG H&L (HRP) ready to use.
Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
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Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling CD10 with ab255609 at 1/1000 dilution, followed by a Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining on proximal convoluted tubules and glomerular epithelial cells of human kidney (PMID:10705818) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a Goat Anti-Rabbit IgG H&L (HRP) ready to use.
Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
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
Certificate of Compliance
文献 (2)
ab255609 被引用在 2 文献中.
- Zeng W et al. CCL18 signaling from tumor-associated macrophages activates fibroblasts to adopt a chemoresistance-inducing phenotype. Oncogene 42:224-237 (2023). PubMed: 36418470
- Yokomizo R et al. Endometrial regeneration with endometrial epithelium: homologous orchestration with endometrial stroma as a feeder. Stem Cell Res Ther 12:130 (2021). PubMed: 33579355