重组Anti-CD86抗体[EPR21962] (ab239075)
![Flow Cytometry - Anti-CD86 antibody [EPR21962] (ab239075)](https://www.abcam.cn/ps/products/239/ab239075/Images/ab239075-326984-anti-cd86-antibody-epr21962-flow-cytometry.jpg "Flow Cytometry - Anti-CD86 antibody [EPR21962] (ab239075)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR21962] to CD86
- Suitable for: WB, ICC/IF, Flow Cyt, IP
- Knockout validated
- Reacts with: Human
概述
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产品名称
Anti-CD86抗体[EPR21962]
参阅全部 CD86 一抗 -
描述
兔单克隆抗体[EPR21962] to CD86 -
宿主
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species Flow Cyt HumanICC/IF HumanIP HumanWB Human
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免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: Ramos, Raji and Daudi whole cell lysates. ICC/IF: Raji and Daudi cells. Flow Cyt: Raji 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: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading... -
纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR21962 -
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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ELISA pair antibody
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Isotype control
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Recombinant Protein
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Related Buffer
应用
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab239075 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 |
|---|---|
| Flow Cyt |
Human
|
| ICC/IF |
Human
|
| IP |
Human
|
| WB |
Human
|
| 应用 | Ab评论 | 说明 |
|---|---|---|
| WB |
1/1000. Detects a band of approximately 80 kDa (predicted molecular weight: 38 kDa).
|
|
| ICC/IF |
1/100.
|
|
| Flow Cyt |
1/500.
|
|
| IP |
1/30.
|
| 说明 |
|---|
|
WB
1/1000. Detects a band of approximately 80 kDa (predicted molecular weight: 38 kDa). |
|
ICC/IF
1/100. |
|
Flow Cyt
1/500. |
|
IP
1/30. |
靶标
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功能
Receptor involved in the costimulatory signal essential for T-lymphocyte proliferation and interleukin-2 production, by binding CD28 or CTLA-4. May play a critical role in the early events of T-cell activation and costimulation of naive T-cells, such as deciding between immunity and anergy that is made by T-cells within 24 hours after activation. Isoform 2 interferes with the formation of CD86 clusters, and thus acts as a negative regulator of T-cell activation. -
组织特异性
Expressed by activated B-lymphocytes and monocytes. -
序列相似性
Contains 1 Ig-like C2-type (immunoglobulin-like) domain.
Contains 1 Ig-like V-type (immunoglobulin-like) domain. -
翻译后修饰
Polyubiquitinated; which is promoted by MARCH8 and results in endocytosis and lysosomal degradation. -
细胞定位
Membrane. - Information by UniProt
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数据库链接
- Entrez Gene: 942 Human
- Omim: 601020 Human
- SwissProt: P42081 Human
- Unigene: 171182 Human
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别名
- Activation B7-2 antigen 3 antibody
- Activation B7-2 antigen antibody
- B-lymphocyte activation antigen B7-2 2 antibody
see all
图片
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Flow Cytometry - Anti-CD86 antibody [EPR21962] (ab239075)
Flow cytometric analysis of Raji (human Burkitt's lymphoma cell line) cell line labeling CD86 with ab239075 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.
Gated on viable cells.
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![Immunocytochemistry/ Immunofluorescence - Anti-CD86 antibody [EPR21962] (ab239075)](https://www.abcam.cn/ps/products/239/ab239075/Images/ab239075-326987-anti-cd86-antibody-epr21962-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-CD86 antibody [EPR21962] (ab239075)Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Daudi (human Burkitt's lymphoma cell line) cells labeling CD86 with ab239075 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining in Daudi cell line is observed.
The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
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![Western blot - Anti-CD86 antibody [EPR21962] (ab239075)](https://www.abcam.cn/ps/products/239/ab239075/Images/ab239075-448036-anti-cd86-antibody-epr21962-western-blot-raji-wt-raji-cd86-knockout.jpg)
Western blot - Anti-CD86 antibody [EPR21962] (ab239075)All lanes : Anti-CD86 antibody [EPR21962] (ab239075) at 1/1000 dilution
Lane 1 : Wild-type Raji cell lysate
Lane 2 : CD86 knockout Raji cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 38 kDa
Observed band size: 70 kDa why is the actual band size different from the predicted?Lanes 1 - 2: Merged signal (red and green). Green - ab239075 observed at 70 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.
ab239075 was shown to react with CD86 in wild-type Raji cells in western blot with loss of signal observed in CD86 knockout sample. Wild-type and CD86 knockout Raji cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab239075 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 hour at room temperature before imaging.
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![Immunocytochemistry/ Immunofluorescence - Anti-CD86 antibody [EPR21962] (ab239075)](https://www.abcam.cn/ps/products/239/ab239075/Images/ab239075-326986-anti-cd86-antibody-epr21962-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-CD86 antibody [EPR21962] (ab239075)Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Raji (human Burkitt's lymphoma cell line) cells labeling CD86 with ab239075 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining in Raji cell line is observed.
The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
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![Immunoprecipitation - Anti-CD86 antibody [EPR21962] (ab239075)](https://www.abcam.cn/ps/products/239/ab239075/Images/ab239075-326985-anti-cd86-antibody-epr21962-immunoprecipitation.jpg)
Immunoprecipitation - Anti-CD86 antibody [EPR21962] (ab239075)CD86 was immunoprecipitated from 0.35 mg of Raji whole (human Burkitt's lymphoma cell line) cell lysate with ab239075 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab239075 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.
Lane 1: Raji whole cell lysate 10 µg (Input).
Lane 2: ab239075 IP in Raji whole lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab239075 in Raji whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes.
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![Western blot - Anti-CD86 antibody [EPR21962] (ab239075)](https://www.abcam.cn/ps/products/239/ab239075/Images/ab239075-326983-anti-cd86-antibody-epr21962-western-blot.jpg)
Western blot - Anti-CD86 antibody [EPR21962] (ab239075)All lanes : Anti-CD86 antibody [EPR21962] (ab239075) at 1/1000 dilution
Lane 1 : Ramos (human Burkitt's lymphoma cell line) whole cell lysate
Lane 2 : Raji (human Burkitt's lymphoma cell line) whole cell lysate
Lane 3 : Daudi (human Burkitt's lymphoma cell line) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 38 kDa
Observed band size: 80 kDa why is the actual band size different from the predicted?Exposure time : Lane 1: 3 minutes; Lanes 2-3: 26 seconds.
Blocking/Dilution buffer: 5% NFDM/TBST.
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![Anti-CD86 antibody [EPR21962] (ab239075)](https://www.abcam.cn/ps/products/239/ab239075/Images/ab239075-6-benefits-of-recombinant-antibodies.png)
Anti-CD86 antibody [EPR21962] (ab239075)
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 (1)
ab239075 被引用在 1 文献中.
- Zhang C et al. miR-30c-5p Reduces Renal Ischemia-Reperfusion Involving Macrophage. Med Sci Monit 25:4362-4369 (2019). PubMed: 31185006