AGAP1 Knockout cell line (MDA-MB-231)

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AGAP1 Knockout cell line (MDA-MB-231)

AGAP1 Knockout cell line (MDA-MB-231) Order

Catalog#YKO-H696

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-H696
Cell line AGAP1 Knockout cell line(MDA-MB-231) Cell Type Human Breast Cancer Cell Line
Morphology Epithelial-like, adherent Passage ratio 1:3~1:4
Culture method 90%L-15+10%FBS
Cryopreservation solution 90%FBS+10%DMSO
Special Note L-15 medium is formulated for growth of cells in CO2 free system. If using CO2 incubator, seal the flask cap tightly to ensure a CO2-free environment

STR Validation

Gene Information

Official symbol
AGAP1
Gene id
116987
Official full symbol
ArfGAP with GTPase domain, ankyrin repeat and PH domain 1
Also known as
AGAP-1, CENTG2, GGAP1, cnt-g2
This gene encodes a member of an ADP-ribosylation factor GTPase-activating protein family involved in membrane trafficking and cytoskeleton dynamics. This gene functions as a direct regulator of the adaptor-related protein complex 3 on endosomes. Multiple transcript variants encoding different isoforms have been found for this gene.

KO strategy

Knockout region: Exon 2 (based on transcript AGAP1-201)
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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