Human ARG1 knockout Hep G2 cell line (ab281603)
製品の概要
-
製品名
Human ARG1 knockout Hep G2 cell line
Liver Arginase ライゼート 製品一覧 -
Parental Cell Line
HepG2 -
Organism
Human -
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
1 -
特記事項
Recommended control: Human wild-type HepG2 cell line (ab275467). 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
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 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.
製品の特性
-
Number of cells
1 x 106 cells/vial, 1 mL -
Adherent /Suspension
Adherent -
Tissue
Liver -
Cell type
epithelial -
Disease
Hepatocellular Carcinoma -
Gender
Male -
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
-
パスウェイ
Nitrogen metabolism; urea cycle; L-ornithine and urea from L-arginine: step 1/1. -
関連疾患
Defects in ARG1 are the cause of argininemia (ARGIN) [MIM:207800]; also known as hyperargininemia. Argininemia is a rare autosomal recessive disorder of the urea cycle. Arginine is elevated in the blood and cerebrospinal fluid, and periodic hyperammonemia occurs. Clinical manifestations include developmental delay, seizures, mental retardation, hypotonia, ataxia, progressive spastic quadriplegia. -
配列類似性
Belongs to the arginase family. -
細胞内局在
Cytoplasm. - Information by UniProt
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ab281603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 |
|---|---|---|
| WB |
Use at an assay dependent concentration.
|
| 特記事項 |
|---|
|
WB
Use at an assay dependent concentration. |
画像
-
Western blot - Human ARG1 knockout Hep G2 cell line (ab281603)All lanes : Anti-Liver Arginase antibody [EPR6671(B)] (ab124917) at 1/1000 dilution
Lane 1 : Wild-type HepG2 cell lysate at 20 µg
Lane 2 : arg1 knockout HepG2 cell lysate at 20 µg
Lanes 3 & 5 : Empty at 0 µg
Lane 4 : Human Liver cell lysate at 5 µg
Lane 6 : A549 cell lysate at 20 µg
Performed under reducing conditions.
Observed band size: 36 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-Liver Arginase antibody [EPR6671(B)] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab124917 was shown to bind specifically to Liver Arginase. A band was observed at 36 kDa in wild-type HepG2 cell lysates with no signal observed at this size in arg1 knockout cell line ab281603 (knockout cell lysate ab282955). Faint band remaining in KO sample at same molecular weight is likely to be an isoform of arg1. To generate this image, wild-type and arg1 knockout HepG2 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 (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
-
Sanger Sequencing - Human ARG1 knockout Hep G2 cell line (ab281603) -
Western blot - Human ARG1 knockout Hep G2 cell line (ab281603)All lanes : Anti-Liver Arginase antibody [EPR6672(B)] (ab133543) at 1/1000 dilution
Lane 1 : Wild-type HepG2 cell lysate at 20 µg
Lane 2 : arg1 knockout HepG2 cell lysate at 20 µg
Lanes 3 & 5 : Empty at 0 µg
Lane 4 : Human Liver cell lysate at 5 µg
Lane 6 : A549 cell lysate at 20 µg
Performed under reducing conditions.
Observed band size: 36 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-Liver Arginase antibody [EPR6672(B)] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab133543 was shown to bind specifically to Liver Arginase. A band was observed at 36 kDa in wild-type HepG2 cell lysates with no signal observed at this size in arg1 knockout cell line ab281603 (knockout cell lysate ab282955). To generate this image, wild-type and arg1 knockout HepG2 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 (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
プロトコール
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.
データシートおよび資料
-
SDS download
-
Datasheet download
参考文献 (0)
ab281603 は論文での使用が確認できていません。