人SCARB2 knockout MCF7 cell line (ab274952)
概述
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产品名称
人SCARB2 knockout MCF7 cell line -
Parental Cell Line
MCF7 -
Organism
Human -
Mutation description
Knockout achieved by CRISPR/Cas9; X = 1 bp insertion; Frameshift: 100% -
Passage number
<20 -
Knockout validation
Next Generation Sequencing (NGS), Western Blot (WB) -
经测试应用
适用于: Next Generation Sequencing, WBmore details -
Biosafety level
1 -
常规说明
Recommended control: Human wild-type MCF7 cell line (ab271144). 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: MEM + 10% FBS + 0.01 mg/ml bovine insulin
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 bath for 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 5-7x104 cells/cm2. 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 5-7x104 cells/cm2 is recommended.
- A partial media change 24 hours prior to subculture may be helpful to encourage growth, if required.
- Cells should be passaged when they have achieved 80-90% confluence.
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
Adherent -
Tissue
Breast -
Cell type
epithelial -
Disease
Adenocarcinoma -
Gender
Female -
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
Acts as a lysosomal receptor for glucosylceramidase (GBA) targeting.
(Microbial infection) Acts as a receptor for enterovirus 71. -
疾病相关
Epilepsy, progressive myoclonic 4, with or without renal failure
Genetic variants in SCARB2 can act as modifier of the phenotypic expression and severity of Gaucher disease. -
序列相似性
Belongs to the CD36 family. -
细胞定位
Lysosome membrane. - Information by UniProt
相关产品
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KO cell lysates
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab274952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 |
|---|---|---|
| Next Generation Sequencing |
Use at an assay dependent concentration.
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| WB |
Use at an assay dependent concentration. Predicted molecular weight: 54 kDa.
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| 说明 |
|---|
|
Next Generation Sequencing
Use at an assay dependent concentration. |
|
WB
Use at an assay dependent concentration. Predicted molecular weight: 54 kDa. |
图片
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Next Generation Sequencing - Human SCARB2 knockout MCF7 cell line (ab274952)1 bp insertion after Leu176 of the WT protein
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Western blot - Human SCARB2 knockout MCF7 cell line (ab274952)All lanes : Anti-LIMPII antibody [EPR12080] (ab176317) at 1/1000 dilution
Lane 1 : Wild-type MCF7 cell lysate
Lane 2 : SCARB2 knockout MCF7 cell lysate
Lane 3 : SH-SY5Y cell lysate
Lane 4 : HEK-293 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 54 kDa
Observed band size: 80 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-LIMPII antibody [EPR12080] 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, ab176317 was shown to bind specifically to LIMPII. A band was observed at 80 kDa in wild-type MCF7 cell lysates with no signal observed at this size in SCARB2 knockout cell line ab274952 (knockout cell lysate ab275010). To generate this image, wild-type and SCARB2 knockout MCF7 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.
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Western blot - Human SCARB2 knockout MCF7 cell line (ab274952)All lanes : Anti-Scavenging Receptor SRB2 antibody [EPR12081] - C-terminal (ab196651) at 1/20000 dilution
Lane 1 : Wild-type MCF7 cell lysate
Lane 2 : SCARB2 knockout MCF7 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 54 kDa
Observed band size: 80 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-Scavenging Receptor SRB2 antibody [EPR12081] - C-terminal staining at 1/20000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab196651 was shown to bind specifically to Scavenging Receptor SRB2. A band was observed at 80 kDa in wild-type MCF7 cell lysates with no signal observed at this size in SCARB2 knockout cell line ab274952 (knockout cell lysate ab275010). To generate this image, wild-type and SCARB2 knockout MCF7 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.
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Next Generation Sequencing - Human SCARB2 knockout MCF7 cell line (ab274952)Knockout achieved by CRISPR/Cas9; X = 1 bp insertion; Frameshift: 100%
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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