重组Anti-CD20抗体[SP32] (ab64088)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [SP32] to CD20
- Suitable for: Flow Cyt (Intra), WB, mIHC, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-CD20抗体[SP32]
参阅全部 CD20 一抗 -
描述
兔单克隆抗体[SP32] to CD20 -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), WB, mIHC, IHC-P, ICC/IFmore details -
种属反应性
与反应: Mouse, Rat, Human
预测可用于: Cow, Dog -
免疫原
Synthetic peptide within Human CD20 aa 250 to the C-terminus. The exact sequence is proprietary.
Database link: P11836 -
阳性对照
- IHC-P: Human tonsil tissue. Mouse and rat spleen tissues. ICC: Ramos cells. Flow Cyt (Intra): WEHI-231 and Ramos cells. WB: Ramos cell lysate.
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常规说明
This product is FOR RESEARCH USE ONLY. For commercial use, please contact partnerships@abcam.com.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
存储溶液
pH: 7.60
Preservative: 0.1% Sodium azide
Constituents: PBS, 1% BSA -
Concentration information loading...
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纯度
Protein A/G purified -
纯化说明
Purified from TCS by protein A/G. -
克隆
单克隆 -
克隆编号
SP32 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Isotype control
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Positive Controls
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab64088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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Flow Cyt (Intra) |
1/40.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB | (1) |
Use at an assay dependent concentration.
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mIHC |
Use at an assay dependent concentration.
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IHC-P | (6) |
1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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ICC/IF | (1) |
1/100.
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说明 |
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Flow Cyt (Intra)
1/40. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
Use at an assay dependent concentration. |
mIHC
Use at an assay dependent concentration. |
IHC-P
1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
ICC/IF
1/100. |
靶标
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功能
This protein may be involved in the regulation of B-cell activation and proliferation. -
组织特异性
Expressed on B-cells. -
疾病相关
Defects in MS4A1 are the cause of immunodeficiency common variable type 5 (CVID5) [MIM:613495]; also called antibody deficiency due to CD20 defect. CVID5 is a primary immunodeficiency characterized by antibody deficiency, hypogammaglobulinemia, recurrent bacterial infections and an inability to mount an antibody response to antigen. The defect results from a failure of B-cell differentiation and impaired secretion of immunoglobulins; the numbers of circulating B cells is usually in the normal range, but can be low. -
序列相似性
Belongs to the MS4A family. -
翻译后修饰
Phosphorylated. Might be functionally regulated by protein kinase(s). -
细胞定位
Membrane. - Information by UniProt
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数据库链接
- Entrez Gene: 485430 Dog
- Entrez Gene: 931 Human
- Entrez Gene: 12482 Mouse
- Entrez Gene: 309217 Rat
- Omim: 112210 Human
- SwissProt: Q3C2E2 Dog
- SwissProt: P11836 Human
- SwissProt: P19437 Mouse
see all -
别名
- APY antibody
- ATOPY antibody
- B lymphocyte antigen CD20 antibody
see all
图片
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All lanes : Anti-CD20 antibody [SP32] (ab64088) at 1/1000 dilution
Lane 1 : Wild-type Raji cell lysate
Lane 2 : MS4A1 knockout Raji cell lysate
Lane 3 : Ramos cell lysate
Lane 4 : A549 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Observed band size: 33 kDa why is the actual band size different from the predicted?Western blot: Anti-MS4A1 antibody [SP32] (ab64088) staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab64088 was shown to bind specifically to MS4A1. A band was observed at 33 kDa in wild-type Raji cell lysates with no signal observed at this size in MS4A1 knockout cell line. To generate this image, wild-type and MS4A1 knockout Raji cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
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This image was generated using ab236434, the same clone, but with a different buffer formulation.
Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Rat spleen tissue labeling Sialoadhesin/CD169, CD3 and CD20 with ab312840 at 1/100 dilution, ab16669 at 1:150 dilution and ab236434 at 1:5000 dilution.
Panel A: merged staining of anti-Sialoadhesin/CD169 (red; Opal™690), anti-CD3 (green; Opal™520) and anti-CD20 (gray; Opal™570) on rat spleen.
Panel B: anti-Sialoadhesin/CD169 stained on macrophages.
Panel C: anti-CD3 stained on T cells.
Panel D: anti-CD20 stained on B cells.The section was incubated in three rounds of staining: in the order of ab312840, ab16669, and ab236434 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.
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This image was generated using ab236434, the same clone, but with a different buffer formulation.
Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse spleen tissue labeling Sialoadhesin/CD169, CD3 and CD20 with ab312840 at 1/100 dilution, ab16669 at 1:150 dilution and ab236434 at 1:5000 dilution.
Panel A: merged staining of anti-Sialoadhesin/CD169 (red; Opal™690), anti-CD3 (green; Opal™520) and anti-CD20 (gray; Opal™570) on rat spleen.
Panel B: anti-Sialoadhesin/CD169 stained on macrophages.
Panel C: anti-CD3 stained on T cells.
Panel D: anti-CD20 stained on B cells.The section was incubated in three rounds of staining: in the order of ab312840, ab16669, and ab236434 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.
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Flow cytometry analysis of Ramos (human Burkitt's lymphoma) labeling CD20 with purified ab64088 at 1/40 dilution (7.78 µg/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as a secondary antibody. Isotype control -Rabbit monoclonal IgG (ab172730) (Black). Unlableled control -Unlabelled cells (blue).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat spleen tissue sections labeling CD20 with ab64088 at 1/100 dilution (3.11 µg/ml). Heat mediated antigen retrieval with citrate buffer (pH 6.0, epitope retrieval solution 1) for 10mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Hematoxylin was used as a counterstain. Positive staining on rat spleen, performed on a Leica Biosystems BOND™ RX instrument.
The section was incubated with ab64088 for 30 mins at room temperature -
Immunocytochemistry/ Immunofluorescence analysis of Ramos (human Burkitt's lymphoma B lymphocyte) cells labeling CD20 with purified ab64088 at 1/100 (3.1 µg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse spleen tissue sections labeling CD20 with ab64088 at 1/100 dilution (3.11 µg/ml). Heat mediated antigen retrieval with citrate buffer (pH 6.0, epitope retrieval solution 1) for 10mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Hematoxylin was used as a counterstain. Positive staining on mouse spleen, performed on a Leica Biosystems BOND™ RX instrument.
The section was incubated with ab64088 for 30 mins at room temperature. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human tonsil tissue sections labeling CD20 with ab64088 at 1/100 dilution (3.11 µg/ml). Heat mediated antigen retrieval with citrate buffer (pH 6.0, epitope retrieval solution 1) for 10mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Hematoxylin was used as a counterstain. Positive staining on human tonsil, performed on a Leica Biosystems BOND™ RX instrument.
The section was incubated with ab64088 for 30 mins at room temperature. -
Immunohistochemical analysis of CD20 expression in formalin fixed, paraffin embedded human tonsil tissue using ab64088 at a 1/100 dilution.
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Flow cytometry analysis of WEHI-231 (mouse lymphoblast B cell lymphoma) labeling CD20 with purified ab64088 at 1/40 dilution (7.78 µg/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as a secondary antibody. Isotype control -Rabbit monoclonal IgG (ab172730) (Black). Unlableled control -Unlabelled cells (blue).
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Anti-CD20 antibody [SP32] (ab64088) at 1/100 dilution + lysate prepared from Ramos cells
Observed band size: 33 kDa why is the actual band size different from the predicted? -
Flow cytometric analysis of rabbit anti-CD20 (SP32) antibody ab64088(1/100) in Jurkat cells (green) compared to negative control of rabbit IgG (blue).
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (11)
ab64088 被引用在 11 文献中.
- Lin C et al. PTEN-induced kinase 1 enhances the reparative effects of bone marrow mesenchymal stromal cells on mice with renal ischaemia/reperfusion-induced acute kidney injury. Hum Cell 35:1650-1670 (2022). PubMed: 35962179
- Mitsui I et al. Chronic Cholecystitis of Dogs: Clinicopathologic Features and Relationship with Liver. Animals (Basel) 11:N/A (2021). IHC-P ; Dog . PubMed: 34828055
- de Melo CVB et al. Splenic Transcriptional Responses in Severe Visceral Leishmaniasis: Impaired Leukocyte Chemotaxis and Cell Cycle Arrest. Front Immunol 12:716314 (2021). PubMed: 34804009
- Yen YT et al. Protein phosphatase 2A inactivation induces microsatellite instability, neoantigen production and immune response. Nat Commun 12:7297 (2021). PubMed: 34911954
- Cybula M et al. Patient-Derived Xenografts of High-Grade Serous Ovarian Cancer Subtype as a Powerful Tool in Pre-Clinical Research. Cancers (Basel) 13:N/A (2021). PubMed: 34944908