Human HTT Knockout HeLa Cell Line

[CAT#: NCL2008ZP554]

Human HTT knockout cell line

Species:: Human

Cell Types:: Other Cells

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Product Overview

Description

This product NCL2008ZP554 is a human HTT gene knockout HeLa cell line which achieved by using CRISPR/Cas9 gene editing technology.

Cell Types

Other Cells

Cell Location

Epithelial

Application Notes

To use as WB control, resuspend the lyophilizate in 50 µL of LDS* Sample Buffer to have a final concentration of 2 mg/ml. For reducing conditions, we recommend a final concentration of 0.1 M DTT.

*Usage of SDS sample buffer is not recommended with these lyophilized lysates.

Mutation Description

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 21

Passage Number

<20

Knockout Validation

Sanger Sequencing, Western Blot (WB)

Relevant Diseases

Huntington's Disease

Research Areas

Neurodegeneration

Species

Human

Properties

Size

1 x 10e6 cells/vial, 1 mL

Formulation

Constituents: 8.7% DMSO, 2% Cellulose, methyl ether

Growth Pattern

Adherent

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

Biosafety Level

Tissue Source

Cervix

Cell Purity

>95%

Cell Viability

~90%

Antibiotic Resistance

Puromycin 1.00µg/ml

Mycoplasma Testing

Yes

Sterility Testing

Sterility testing was performed in accordance with USP and EP regulations. All of our sterility testing is performed in an isolator or clean room environments. The cell line has been screened using the membrane filtration testing methods to confirm the absence of aerobic, anaerobic and fungi microorganisms.

Genetic Stability Testing

Cell genetic stability study was perfomed under ICH guidelines. We provide guidance on the appropriate testing program upon your requirements.

Shipping

Dry ice

Storage

Shipped on Dry Ice. Store in liquid nitrogen.

Handling Advice

Upon arrival, the sample vial should be stored in liquid nitrogen vapor phase instead of -80°C. Storage at -80°C will cause loss of vitality.

1. Thaw the vial in a 37°C water bath for about 1-2 minutes.
2. Transfer the cell suspension to a 15 mL conical tube containing pre-warmed 5 mL complete medium DMEM + 10% FBS, and rotate 125 x g for about 5 minutes at room temperature.
3. Resuspend the cell pellet with 1 mL of pre-warmed complete medium DMEM + 10% FBS, and distribute it into a 25 cm2 culture flask, which contains 10 mL of pre-warmed complete medium DMEM + 10% FBS.
4. Incubate the culture with 5% CO2 in a 37°C incubator.
5. The recommended subculture ratio is 1:4-1:6. When cells grow at 80-90% confluence and divide, the cells should be passaged.

Research Use Only

For research use only, not for diagnostic or therapeutic use.

Warnings

Store in gas phase of liquid nitrogen immediately upon receipt. This product is stable for 6 months when stored as directed.

Quality Control

Mycoplasma was tested for sterility, post-freezing viability, and short-term repeat (STR) analysis for cell line identification. Cytochrome oxidase I (COI) was analyzed for cell line species identification.

Target Details

Target

Huntingtin

Official Name

HTT

Full Name

huntingtin gene

Alternative Names

HTT; HD; IT15; huntingtin; LOMARS

Gene ID

3064(Human); 15194(Mouse); 29424(Rat)

Uniprot ID

P42858(Human); P42859(Mouse); P51111(Rat)

Publications

Publications (0)

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