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AB266515

Human APEH (APH) knockout HEK-293T cell line

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APEH KO cell line available to order. KO validated. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 4. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.

대체 명칭 보기

DNF15S2, APEH, Oxidized protein hydrolase, APH, Acylamino-acid-releasing enzyme, D3F15S2, Acyl-peptide hydrolase, OPH, Acylaminoacyl-peptidase, D3S48E, AARE

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Sanger Sequencing - Human APEH (APH) knockout HEK-293T cell line (AB266515)
  • Sanger seq

Unknown

Sanger Sequencing - Human APEH (APH) knockout HEK-293T cell line (AB266515)

Homozygous : 1 bp insertion in exon4

주요 정보

세포 유형

HEK-293T

Species or organism

Human

조직

Kidney

제형

Liquid

form

Knockout 검증

Sanger Sequencing

Mutation 정보

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 4

제품 세부 정보

We will provide viable cells that proliferate on revival.

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.

제품 구성

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특성 및 보관 정보

유전자명
APEH
유전자 편집 종류
Knockout
유전자 편집 방식
CRISPR technology
Knockout 검증
Sanger Sequencing
Zygosity
Homozygous
배송 시 보관 조건
Dry Ice
적절한 단기 보관 조건
-196°C
적절한 장기 보관 조건
-196°C

추가 정보

This supplementary information is collated from multiple sources and compiled automatically.

The target protein APH also known as Aminoglycoside Phosphotransferase or APH(2") serves a critical role in antibiotic resistance mechanisms by phosphorylating aminoglycoside antibiotics leading to inactivation. APH with a mass of approximately 31 kDa can be found in bacteria that are often resistant to several antibiotics. Although mainly located in prokaryotes certain modified forms also show expression in eukaryotic systems for research purposes.
Biological function summary

APH plays a significant role in cell survival by protecting bacteria from antibiotic stress a process necessary for maintaining membrane integrity under antibiotic pressure. This enzyme is not a part of a larger protein complex but exhibits highly specific interactions with aminoglycosides. It commonly associates with resistance mechanisms allowing bacteria to survive in otherwise lethal environments.

Pathways

APH significantly impacts aminoglycoside metabolic pathways and contributes to the overarching antibiotic resistance pathway. This phosphotransferase action is often coordinated alongside AAC (Aminoglycoside N-acetyltransferase) and ANT (Aminoglycoside O-nucleotidyltransferase) proteins. They collectively enable bacteria survival by modifying antibiotics preventing their effective binding to bacterial ribosomal targets.

APH presents a considerable threat due to its association with multidrug-resistant bacterial infections and urinary tract infections. The protein's role in resistance complicates treatment protocols and is integrated into bacterial adaptation strategies. Alongside proteins like AAC and ANT APH contributes to bacterial resilience making it a target of interest in studies focused on developing novel antibacterial treatments.

품질 관리

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

초기 취급 지침

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. <p>1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.<br>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.<br>3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.<br>4. Incubate the culture at 37°C incubator with 5% CO<sub>2</sub>. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.<br>5. Once confluent passage into an appropriate flask at a density of 2x10<sup>4</sup> cells/cm<sup>2</sup>. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.</p>

Subculture 가이드라인
  • All seeding densities should be based on cell counts gained by established methods.
  • A guide seeding density of 2x104 cells/cm2 is recommended.
  • Cells should be passaged when they have achieved 80-90% confluence.
배양 배지

DMEM (High Glucose) + 10% FBS

동결보존 배지

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

제품 프로토콜

Product promise

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