ISG15 Knockout cell line (THP-1)

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ISG15 Knockout cell line (THP-1)

ISG15 Knockout cell line (THP-1) Order

Catalog#YKO-H722

Price (USD)-

Size 1*10^6

Instruction

Exclusive Red Cotton system(en.rc-crispr.com)was used to design the sgRNA fragment knockout strategy. Two sgRNAs were constructed into CRISPR-U™ plasmid which will be subsequentially transfected into cells by electroporation/virus method. And the single-cell clones were isolated by the limited dilution method and the Monoclone Genotype Validation Kit (Cat: YK-MV-1000) was used for cell lysis and PCR amplification on the single-cell clones, and then the genotype was verified by Sanger sequencing. Upon passing the final validation, the positive clones were expanded and cryopreserved.

Cell Info

Catalog YKO-H722
Cell line ISG15 Knockout cell line(THP-1) Cell Type Human Ovarian Granulosa Cell Line
Morphology Monocyte, suspension Passage ratio Control the cell density at 2x10^5~4.0x10^5 cells/ml during passaging, and carry out the passage when the cells grow to 8x10^5~1.0x10^6 cells/ml.
Culture method 90%RPMI-1640+10%FBS
Cryopreservation solution 90%FBS+10%DMSO
Special Note The cell culture density needs to be strictly controlled. The cell viability after thawing is relatively low, it is recommended to use Australia-sourced FBS for thawing or increase the FBS ratio to 20%. When the cells grow normally and can be passaged after 2 passages, the cells can be cultured using the normal culture method.

STR Validation

Gene Information

Official symbol
ISG15
Gene id
9636
Official full symbol
ISG15 ubiquitin like modifier
Also known as
G1P2, IFI15, IMD38, IP17, UCRP, hUCRP
The protein encoded by this gene is a ubiquitin-like protein that is conjugated to intracellular target proteins upon activation by interferon-alpha and interferon-beta. Several functions have been ascribed to the encoded protein, including chemotactic activity towards neutrophils, direction of ligated target proteins to intermediate filaments, cell-to-cell signaling, and antiviral activity during viral infections. While conjugates of this protein have been found to be noncovalently attached to intermediate filaments, this protein is sometimes secreted.

KO strategy

Knockout region: Exon 2 (based on transcript ISG15-203)
Validation: PCR+Sanger Sequencing

Method

Exclusive Red Cotton system(en.rc-crispr.com)was used to design the sgRNA fragment knockout strategy. Two sgRNAs were constructed into CRISPR-U™ plasmid which will be subsequentially transfected into cells by electroporation/virus method. And the single-cell clones were isolated by the limited dilution method and the Monoclone Genotype Validation Kit (Cat: YK-MV-1000) was used for cell lysis and PCR amplification on the single-cell clones, and then the genotype was verified by Sanger sequencing. Upon passing the final validation, the positive clones were expanded and cryopreserved.

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