人TCF12 knockout Jurkat cell line (ab274931)
概述
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产品名称
人TCF12 knockout Jurkat cell line
参阅全部 TCF12 细胞裂解液 -
Parental Cell Line
Jurkat -
Organism
Human -
Mutation description
Knockout achieved by CRISPR/Cas9; X =5 bp insertion (allele 1) and 4 bp insertion (allele 2) after Gly47 of the WT protein Frameshift: 100% -
Passage number
<20 -
Knockout validation
Western Blot (WB) -
经测试应用
适用于: WBmore details -
Biosafety level
1 -
常规说明
Recommended control: Human wild-type Jurkat cell line (ab271143). 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.
Cryopreservation cell medium: Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
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 1x105 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.Subculture guidelines:
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 1x105 cells/mL is recommended.
- Do not allow the cell density to exceed 3x106.
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.
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
Blood -
Cell type
T cell lymphoblast-like -
Disease
Non-Hodgkin 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
Binds specifically to oligomers of E-box motifs. May play important roles during development of the nervous system as well as in other organ systems. -
组织特异性
Expressed in several tissues and cell types including skeletal muscle, thymus, and a B-cell line. -
序列相似性
Contains 1 basic helix-loop-helix (bHLH) domain. -
结构域
the 9aaTAD motif is a transactivation domain present in a large number of yeast and animal transcription factors. -
翻译后修饰
Phosphorylated upon DNA damage, probably by ATM or ATR. -
细胞定位
Nucleus. - Information by UniProt
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab274931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: 72 kDa.
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| 说明 |
|---|
|
WB
Use at an assay dependent concentration. Predicted molecular weight: 72 kDa. |
图片
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Next Generation Sequencing - Human TCF12 knockout Jurkat cell line (ab274931)5 bp insertion (allele 1) and 4 bp insertion (allele 2) after Gly47 of the WT protein
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Western blot - Human TCF12 knockout Jurkat cell line (ab274931)All lanes : Anti-TCF12 antibody [7B3] (ab243149) at 0.394 µg/ml
Lane 1 : Wild-type Jurkat cell lysate
Lane 2 : TCF12 CRISPR-Cas9 edited Jurkat cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : MOLT-4 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 72 kDa
Observed band size: 85 kDa why is the actual band size different from the predicted?Western blot: Anti-TCF12 antibody [7B3] staining at 0.394 ug/ml, shown in black; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab243149 was shown to bind specifically to TCF12. A band was observed at 85 kDa in wild-type Jurkat cell lysates with no signal observed at this size in TCF12 CRISPR-Cas9 edited cell line ab274931 (CRISPR-Cas9 edited cell lysate ab274989). The band observed in the CRISPR-Cas9 edited lysate lane below 85 kDa is likely to represent a truncated form of TCF12. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and TCF12 CRISPR-Cas9 edited Jurkat 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 before development with Optiblot (ECL reagent ab133456) and imaged with 20 minutes exposure time. Secondary antibodies used were HRP conjugated Goat anti-Rat (H+L) and Goat anti-Rabbit IgG H&L 680RD 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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