人CD86 knockout Raji cell裂解物(ab273812)
概述
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产品名称
人CD86 knockout Raji cell裂解物
参阅全部 CD86 试剂盒 -
产品概述
Knockout cell lysate achieved by CRISPR/Cas9.
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Parental Cell Line
Raji -
Organism
Human -
Mutation description
Knockout achieved by CRISPR/Cas9; X = 1 bp insertion; Frameshift: 99% -
Passage number
<20 -
Knockout validation
Next Generation Sequencing (NGS), Western Blot (WB) -
Reconstitution notes
To use as WB control, resuspend the lyophilizate in 50 µL of LDS* Sample Buffer to have a final concentration of 2 mg/ml. For reducing conditions, we recommend a final concentration of 0.1 M DTT.*Usage of SDS sample buffer is not recommended with these lyophilized lysates.
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说明
Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version - found here. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
See here for more information on knockout cell lysates.Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
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经测试应用
适用于: WBmore details
性能
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存放说明
Store at -80°C. Please refer to protocols. -
组件 1 kit ab280662 - Human CD86 knockout Raji cell lysate 1 x 100µg ab269600 - Human wild-type Raji cell lysate 1 x 100µg -
研究领域
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Cell type
Burkitt's lymphoma -
Disease
Lymphoma -
Gender
Male
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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别名
- Activation B7-2 antigen
- Activation B7-2 antigen 3
- B-lymphocyte activation antigen B7-2
see all
相关产品
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KO cell lines
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Related Products
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应用
The Abpromise guarantee
Abpromise™承诺保证使用ab273812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 |
Use at an assay dependent concentration.
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说明 |
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WB
Use at an assay dependent concentration. |
图片
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Knockout achieved by CRISPR/Cas9; X = 1 bp insertion; Frameshift: 99%
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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
Lane 3: Ramos whole cell lysate
Lane 4: Human liver cell lysateLanes 1 - 4: Merged signal (red and green). Green - ab220188 observed at 70 kDa. Red - loading control ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55 kDa.
ab220188 was shown to react with CD86 in wild-type Raji cells in Western blot with loss of signal observed in CD86 knockout cell line ab273812 (knockout cell lysate ab273858). Wild-type Raji and CD86 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab220188 and ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4 °C at 0.5 µg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
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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
Lanes 1 - 2: Merged signal (red and green). Green - ab269587 observed at 70 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.
ab269587 was shown to react with CD86 in wild-type Raji cells in western blot with loss of signal observed in CD86 knockout cell line ab273858 (knockout cell lysate ab273812). 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 ab269587 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. -
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
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 cell line ab273858 (knockout cell lysate ab273812). 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.
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 (0)
ab273812 尚未被引用在任何文献中。