重组Anti-CD86抗体[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
Related conjugates and formulations
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 -
经测试应用
适用于: WB, ICC/IF, Flow Cyt, IPmore details
不适用于: IHC-P -
种属反应性
与反应: Human -
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, Human monocyte-derived dendritic 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...
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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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KO cell lines
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KO cell lysates
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Recombinant Protein
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Related Buffer
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab239075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 80 kDa (predicted molecular weight: 38 kDa).
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ICC/IF |
1/100.
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Flow Cyt |
1/500.
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IP |
1/30.
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说明 |
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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 staining of human monocyte-derived dendritic cells (top) or human monocyte-derived dendritic cells treated with 1μg/mL lipopolysaccharide (LPS) for 24h (bottom), with ab239075 (right) or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (left). Monocyte-derived dendritic cells were incubated for 30 min at 4°C in 1x PBS containing 10 µg/ml human IgG and 10 % normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody ab239075 or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (1x 106 in 100μl at 0.04 μg/ml (1/56500)) for 30min on ice. The cells were simultaneously stained with CD209.
The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 4°C
Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. Events were gated on viable cells.
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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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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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All lanes : Anti-CD86 antibody [EPR21962] (ab239075) at 1/1000 dilution
Lane 1 : Wild-type Raji (Human Burkitt's lymphoma cell line) whole cell lysate
Lane 2 : CD86 knockout Raji (Human Burkitt's lymphoma cell line) whole 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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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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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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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.
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 (14)
ab239075 被引用在 14 文献中.
- Andreuzzi E et al. Colorectal cancer development is affected by the ECM molecule EMILIN-2 hinging on macrophage polarization via the TLR-4/MyD88 pathway. J Exp Clin Cancer Res 41:60 (2022). PubMed: 35148799
- Luo L et al. LPS Activated Macrophages Induced Hepatocyte Pyroptosis via P2X7R Activation of NLRP3 in Mice. Iran J Immunol 19:4 (2022). PubMed: 35293345
- Rogal J et al. Autologous Human Immunocompetent White Adipose Tissue-on-Chip. Adv Sci (Weinh) 9:e2104451 (2022). PubMed: 35466539
- Zhu Y et al. Flavokawain B Weakens Gastric Cancer Progression via the TGF-β1/SMAD4 Pathway and Attenuates M2 Macrophage Polarization. J Immunol Res 2022:4903333 (2022). PubMed: 35879950
- Chen L et al. Construction and analysis of competing endogenous RNA network and patterns of immune infiltration in abdominal aortic aneurysm. Front Cardiovasc Med 9:955838 (2022). PubMed: 35990982