Human CPS1 knockout HeLa cell line (ab261809)
製品の概要
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製品名
Human CPS1 knockout HeLa cell line -
Parental Cell Line
HeLa -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 15 and Insertion of the selection cassette in exon 15 -
Passage number
<20 -
Knockout validation
Sanger Sequencing, Western Blot (WB) -
アプリケーション
適用あり: WBmore details -
Biosafety level
2 -
特記事項
Recommended control: Human wild-type HeLa cell line (ab255928). 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: DMEM (High Glucose) + 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 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 2x104 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 2x104 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.
製品の特性
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Number of cells
1 x 106 cells/vial, 1 mL -
Adherent /Suspension
Adherent -
Tissue
Cervix -
Cell type
epithelial -
Disease
Adenocarcinoma -
Gender
Female -
STR Analysis
Amelogenin X D5S818: 11, 12 D13S317: 12, 13.3 D7S820: 8, 12 D16S539: 9, 10 vWA: 16, 18 TH01: 7 TPOX: 8, 12 CSF1PO: 9, 10 -
Mycoplasma free
Yes -
保存方法
Shipped on Dry Ice. Store in liquid nitrogen. -
バッファー
Constituents: 8.7% Dimethylsulfoxide, 2% Cellulose, methyl ether -
研究分野
ターゲット情報
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機能
Involved in the urea cycle of ureotelic animals where the enzyme plays an important role in removing excess ammonia from the cell. -
組織特異性
Primarily in the liver and small intestine. -
関連疾患
Defects in CPS1 are the cause of carbamoyl phosphate synthetase 1 deficiency (CPS1D) [MIM:237300]. CPS1D is an autosomal recessive disorder of the urea cycle causing hyperammonemia. Clinical features include protein intolerance, intermittent ataxia, seizures, lethargy, developmental delay and mental retardation.
Note=Genetic variations in CPS1 influence the availability of precursors for nitric oxide (NO) synthesis and play a role in clinical situations where endogenous NO production is critically important, such as neonatal pulmonary hypertension, increased pulmonary artery pressure following surgical repair of congenital heart defects or hepatovenocclusive disease following bone marrow transplantation. Infants with neonatal pulmonary hypertension homozygous for Thr-1406 have lower L-arginine concentrations than neonates homozygous for Asn-1406. -
配列類似性
Contains 2 ATP-grasp domains.
Contains 1 glutamine amidotransferase type-1 domain. -
ドメイン
The type-1 glutamine amidotransferase domain is defective. -
細胞内局在
Mitochondrion. - Information by UniProt
関連製品
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KO cell lysates
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Related Products
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab261809の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 165 kDa.
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特記事項 |
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WB
Use at an assay dependent concentration. Predicted molecular weight: 165 kDa. |
画像
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All lanes : Anti-CPS1 antibody [EPR7493-29] (ab155083) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : CPS1 knockout HeLa cell lysate
Lane 3 : Human liver tissue lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 165 kDa
Observed band size: 165 kDaLanes 1-3: Merged signal (red and green). Green - ab155083 observed at 165 kDa. Red - loading control, ab8245 observed at 37 kDa.
ab155083 Anti-CPS1 antibody [EPR7493-29] was shown to specifically react with CPS1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261809 (knockout cell lysate ab257121) was used. Wild-type and CPS1 knockout samples were subjected to SDS-PAGE. ab155083 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-CPS1 antibody [EPR7493-3] (ab129076) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : CPS1 knockout HeLa cell lysate
Lane 3 : Human liver tissue lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 165 kDa
Observed band size: 165 kDaLanes 1-3: Merged signal (red and green). Green - ab129076 observed at 165 kDa. Red - loading control, ab8245 observed at 37 kDa.
ab129076 Anti-CPS1 antibody [EPR7493-3] was shown to specifically react with CPS1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261809 (knockout cell lysate ab257121) was used. Wild-type and CPS1 knockout samples were subjected to SDS-PAGE. ab129076 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
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Allele-1: 1 bp insertion in exon 15.
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Allele-2: Insertion of the selection cassette in exon 15.
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (0)
ab261809 は論文での使用が確認できていません。