人CD27A knockout Raji cell裂解物(ab281360)
概述
-
产品名称
人CD27A knockout Raji cell裂解物
参阅全部 CD27 试剂盒 -
产品概述
Knockout cell lysate achieved by CRISPR/Cas9.
-
Parental Cell Line
Raji -
Organism
Human -
Mutation description
Knockout achieved by CRISPR/Cas9; X = 10 bp deletion; Frameshift: 100% -
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.
-
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 and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
-
经测试应用
适用于: WBmore details
性能
-
存放说明
Store at -80°C. Please refer to protocols. -
组件 1 kit ab281412 - Human CD27A knockout Raji cell lysate 1 x 100µg ab269600 - Human wild-type Raji cell lysate 1 x 100µg -
研究领域
-
Cell type
Burkitt's lymphoma -
Disease
Lymphoma -
Gender
Male
靶标
-
功能
Receptor for CD70/CD27L. May play a role in survival of activated T-cells. May play a role in apoptosis through association with SIVA1. -
组织特异性
Found in most T-lymphocytes. -
序列相似性
Contains 3 TNFR-Cys repeats. -
翻译后修饰
Phosphorylated and O-glycosylated. -
细胞定位
Membrane. - Information by UniProt
-
别名
- CD 27
- CD27
- CD27 antigen
see all
相关产品
-
KO cell lines
-
Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab281360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 |
|---|---|---|
| WB |
Use at an assay dependent concentration.
|
| 说明 |
|---|
|
WB
Use at an assay dependent concentration. |
图片
-
Next Generation Sequencing - Human CD27A knockout Raji cell lysate (ab281360)Knockout achieved by CRISPR/Cas9; X = 10 bp deletion; Frameshift: 100%
-
Western blot - Human CD27A knockout Raji cell lysateLane 1: Wild-type Raji cell lysate (20 ug)
Lane 2: CD27A knockout Raji cell lysate (20 ug)
Lane 3: Ramos cell lysate (20 ug)
Lane 4: Human brain cell lysate (20 ug)ab133761 was shown to react with CD27 in wild-type Raji cells in western blot. Loss of signal was observed when CD27 knockout cell line ab274910 (knockout cell lysate ab281360) was used. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab133761 overnight at 4 °C at a 1 in 1000 dilution. Blots were incubated with HRP conjugated Goat anti-Rabbit (H+L) secondary antibody at 1 in 5000 for 1 hour at room temperature before development with Optiblot ECL reagent (ab133456) and imaging.
-
Western blot - Human CD27A knockout Raji cell lysate (ab281360)Lane 1: Wild-type Raji (Human Burkitt's lymphoma cell line) whole cell lysate 20 ug
Lane 2: CD27 knockout Raji (Human Burkitt's lymphoma cell line) whole cell lysate 20 ug
Lane 3: Ramos (Human Burkitt's lymphoma cell line) whole cell lysate 20 ug
Lane 4: Human brain tissue lysate 20 ug
Lanes 1 - 4: Merged signal (red and green). Green - ab131254 observed at 35 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.
ab131254 was shown to react with CD27 in wild-type Raji cells in western blot with loss of signal observed in CD27 knockout cell line ab274910 (knockout cell lysate ab281360). Wild-type and CD27 knockout Raji cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab131254 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
-
SDS download
-
Datasheet download
文献 (0)
ab281360 尚未被引用在任何文献中。