人CD791 knockout Raji cell line (ab274911)
概述
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产品名称
人CD791 knockout Raji cell line
参阅全部 CD79a 细胞裂解液 -
Parental Cell Line
Raji -
Organism
Human -
Passage number
<20 -
Knockout validation
Next Generation Sequencing (NGS), Western Blot (WB) -
经测试应用
适用于: WBmore details -
Biosafety level
2 -
常规说明
Recommended control: Human wild-type Raji cell line (ab271145)
Cryopreservation cell medium: Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
Culture medium: RPMI + 10% FBS
Initial handling guidelines: Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water for bath approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method. Based on cell count, seed cells in an appropriate cell culture flask at a density of 4x105 cells/mL. Seeding density is given as a guide only and should be scaled to align with individual lab schedules.
4. Incubate the culture at 37°C incubator with 5% CO2. Cultures should be monitored daily.Initial handling guidelines:
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 4x105 cells/mL is recommended.
- A maximum of 3x106 viable cells/ml is obtainable.
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.
We will provide viable cells that proliferate on revival.
性能
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Number of cells
1 x 106 cells/vial, 1 mL -
Adherent /Suspension
Suspension -
Tissue
Lymphatic -
Cell type
Burkitt's lymphoma -
Disease
Lymphoma -
Gender
Male -
Mycoplasma free
Yes -
存放说明
Shipped on Dry Ice. Store in liquid nitrogen. -
存储溶液
Constituents: 8.7% Dimethylsulfoxide, 2% Cellulose, methyl ether -
研究领域
靶标
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功能
Required in cooperation with CD79B for initiation of the signal transduction cascade activated by binding of antigen to the B-cell antigen receptor complex (BCR) which leads to internalization of the complex, trafficking to late endosomes and antigen presentation. Also required for BCR surface expression and for efficient differentiation of pro- and pre-B-cells. Stimulates SYK autophosphorylation and activation. Binds to BLNK, bringing BLNK into proximity with SYK and allowing SYK to phosphorylate BLNK. Also interacts with and increases activity of some Src-family tyrosine kinases. Represses BCR signaling during development of immature B cells. -
组织特异性
B-cells. -
疾病相关
Defects in CD79A are the cause of agammaglobulinemia type 3 (AGM3) [MIM:613501]. It is a primary immunodeficiency characterized by profoundly low or absent serum antibodies and low or absent circulating B cells due to an early block of B-cell development. Affected individuals develop severe infections in the first years of life. Note=Two different mutations, one at the splice donor site of intron 2 and the other at the splice acceptor site for exon 3, have been identified. Both mutations give rise to a truncated protein. -
序列相似性
Contains 1 Ig-like C2-type (immunoglobulin-like) domain.
Contains 1 ITAM domain. -
翻译后修饰
Phosphorylated on tyrosine, serine and threonine residues upon B-cell activation. Phosphorylation of tyrosine residues by Src-family kinases is an early and essential feature of the BCR signaling cascade. The phosphorylated tyrosines serve as docking sites for SH2-domain containing kinases, leading to their activation which in turn leads to phosphorylation of downstream targets. Phosphorylation of serine and threonine residues may prevent subsequent tyrosine phosphorylation. -
细胞定位
Cell membrane. Following antigen binding, the BCR has been shown to translocate from detergent-soluble regions of the cell membrane to lipid rafts although signal transduction through the complex can also occur outside lipid rafts. - Information by UniProt
相关产品
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KO cell lysates
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab274911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. Predicted molecular weight: 25 kDa.
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| 说明 |
|---|
|
WB
Use at an assay dependent concentration. Predicted molecular weight: 25 kDa. |
图片
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Next Generation Sequencing - Human CD791 knockout Raji cell line (ab274911)14 bp deletion after Ser58 of the WT protein
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Western blot - Human CD791 knockout Raji cell line (ab274911)All lanes : Anti-CD79a antibody [EPR3619] (ab133483) at 1/1000 dilution
Lane 1 : Wild-type Raji cell lysate
Lane 2 : CD79A knockout Raji cell lysate
Lane 3 : Daudi cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 25 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-CD79a antibody [EPR3619] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab133483 was shown to bind specifically to CD79a. A band was observed at 50 kDa in wild-type Raji cell lysates with no signal observed at this size in CD79A knockout cell line ab274911 (knockout cell lysate ab281361). To generate this image, wild-type and CD79A 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 5 % 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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Western blot - Human CD791 knockout Raji cell line (ab274911)All lanes : Anti-CD79a antibody [EP3618] (ab79414) at 1/5000 dilution
Lane 1 : Wild-type Raji cell lysate
Lane 2 : CD79A knockout Raji cell lysate
Lane 3 : Daudi cell lysate
Lane 4 : HAP1 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 25 kDa
Observed band size: 40-50 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-CD79a antibody [EP3618] staining at 1/5000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab79414 was shown to bind specifically to CD79a. A band was observed at 40-50 kDa in wild-type Raji cell lysates with no signal observed at this size in CD79A knockout cell line ab274911 (knockout cell lysate ab281361). To generate this image, wild-type and CD79A 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 (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (0)
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