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AB261833

Human PDCD4 knockout HeLa cell line

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PDCD4 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 7. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
2 이미지
Western blot - Human PDCD4 knockout HeLa cell line (AB261833)
  • WB

Lab

Western blot - Human PDCD4 knockout HeLa cell line (AB261833)

Lanes 1- 2 : Merged signal (red and green). Green - ab80590 observed at 51 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab80590 was shown to react with PDCD4 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261833 (knockout cell lysate ab257278) was used. Wild-type HeLa and PDCD4 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab80590 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 20000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-PDCD4 antibody [EPR3431] (<a href='/ko/products/primary-antibodies/pdcd4-antibody-epr3431-ab80590'>ab80590</a>) at 1/20000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human PDCD4 knockout HeLa cell lysate (<a href='/ko/products/cell-lysates/human-pdcd4-knockout-hela-cell-lysate-ab257278'>ab257278</a>) at 20 µg

Predicted band size: 51 kDa

Observed band size: 51 kDa

false

Sanger Sequencing - Human PDCD4 knockout HeLa cell line (AB261833)
  • Sanger seq

Unknown

Sanger Sequencing - Human PDCD4 knockout HeLa cell line (AB261833)

Homozygous : 1 bp insertion in exon 7.

주요 정보

세포 유형

HeLa

Species or organism

Human

조직

Cervix

제형

Liquid

form

Knockout 검증

Sanger Sequencing,Western blot

Mutation 정보

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 7

Antibiotic resistance

Puromycin 1µg/mL

질병

Adenocarcinoma

Reactivity 정보

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

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
PDCD4
유전자 편집 종류
Knockout
유전자 편집 방식
CRISPR technology
Knockout 검증
Sanger Sequencing, Western blot
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 PDCD4 protein also known as Programmed Cell Death 4 is a multifunctional tumor suppressor. With a molecular mass of approximately 64 kDa it plays an important role in regulating gene expression and inhibiting cellular transformation. PDCD4 is expressed in various tissues including the lung kidney and heart where it modulates different cellular pathways. Its ability to bind RNA and interact with eukaryotic initiation factor 4A (eIF4A) highlights its important role in translation regulation.
Biological function summary

PDCD4 inhibits translation by binding to the eIF4A RNA helicase blocking the translation initiation complex. This action stabilizes mRNA and reduces protein synthesis impacting processes such as cell growth and proliferation. Although PDCD4 functions mainly as a monomer it can also interact with other proteins to form complexes that enhance its regulatory activity. More specifically its role in controlling AP-1 transcription factor activity steps into the spotlight in influencing cellular responses.

Pathways

PDCD4 primarily interacts with signaling mechanisms related to apoptosis and inflammation. The protein is integral in the mTOR signaling pathway which governs cell growth in response to nutrient availability. PDCD4 also impacts the PI3K/AKT pathway linking its activity to cellular survival and metabolism. Through these pathways PDCD4 associates with proteins such as p53 and NF-kB connecting it to broader cellular and molecular networks involved in maintaining cellular integrity.

PDCD4 is significantly implicated in cancer progression and inflammation-related conditions. Its downregulation or loss frequently occurs in several cancers including breast and lung cancer leading to unchecked cell proliferation and tumor growth. PDCD4's interaction with proteins like c-Myc further demonstrates its importance in oncogenic pathways. Additionally its role in inflammation positions it as a critical factor in disorders such as Crohn's disease where abnormal inflammatory responses are prominent.

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

타겟 정보

See full target information PDCD4

대체 명칭 보기

PDCD4, Programmed cell death protein 4, Nuclear antigen H731-like, Protein 197/15a, Neoplastic transformation inhibitor protein, H731

Product promise

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