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AB257278

Human PDCD4 knockout HeLa cell lysate

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PDCD4 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 7.

View Alternative Names

DUG, Death up-regulated gene protein, H731, MA 3, MGC33046, MGC33047, Neoplastic transformation inhibitor, Neoplastic transformation inhibitor protein, Nuclear antigen H731, Nuclear antigen H731 like protein, Nuclear antigen H731-like, PDCD4_HUMAN, Programmed cell death 4, Programmed cell death protein 4, Protein 197/15a, Protein MA-3, Tis, Topoisomerase-inhibitor suppressed protein, programmed cell death 4 (neoplastic transformation inhibitor)

3 Images
Western blot - Human PDCD4 knockout HeLa cell lysate (AB257278)
  • WB

Lab

Western blot - Human PDCD4 knockout HeLa cell lysate (AB257278)

Lane 1 : Wild-type HeLa cell lysate (20µg)

Lane 2 : PDCD4 knockout HeLa cell lysate (20µg)

Lanes 1- 2 : Merged signal (red and green). Green - ab80590 observed at 51 kDa. Red - loading control ab8245 observed at 37 kDa.

ab80590 Anti-PDCD4 antibody [EPR3431] was shown to specifically 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 and PDCD4 knockout samples 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) were incubated overnight at 4°C at 1 in 20000 dilution and 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='/en-us/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:

PDCD4 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human PDCD4 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-pdcd4-knockout-hela-cell-line-ab261833'>ab261833</a>)

Predicted band size: 51 kDa

Observed band size: 51 kDa

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Western blot - Human PDCD4 knockout HeLa cell lysate (AB257278)
  • WB

Lab

Western blot - Human PDCD4 knockout HeLa cell lysate (AB257278)

Lane 1 : Wild-type HeLa cell lysate (20µg)

Lane 2 : PDCD4 knockout HeLa cell lysate (20µg)

Lanes 1- 2 : Merged signal (red and green). Green - ab79405 observed at 51 kDa. Red - loading control ab8245 observed at 37 kDa.

ab79405 Anti-PDCD4 antibody [EPR3432] was shown to specifically 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 and PDCD4 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab79405 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 5000 dilution and 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 [EPR3432] (<a href='/en-us/products/primary-antibodies/pdcd4-antibody-epr3432-ab79405'>ab79405</a>) at 1/5000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

PDCD4 knockout HeLa cell lysate at 20 µg

Predicted band size: 51 kDa

Observed band size: 51 kDa

false

Sanger Sequencing - Human PDCD4 knockout HeLa cell lysate (AB257278)
  • Sanger seq

Unknown

Sanger Sequencing - Human PDCD4 knockout HeLa cell lysate (AB257278)

Homozygous : 1 bp insertion in exon 7

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Knockout validation

Sanger Sequencing,Western blot

Mutation description

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

Disease

Adenoma

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Reactivity data

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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

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.

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What's included?

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Properties and storage information

Gene name
PDCD4
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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Supplementary information

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.
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Quality control

STR analysis

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

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Complementary Products

Primary Antibodies

AB80590

Anti-PDCD4 antibody [EPR3431]

BOND RX™ Validated|RabMAb|Recombinant|KO Validated

Flow Cytometry (Intracellular) - Anti-PDCD4 antibody [EPR3431] (AB80590)

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Proteins & Peptides

AB172822

Recombinant Human PDCD4 protein

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Product promise

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For full details, please see our Terms & Conditions

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