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AB257017

Human PARP1 knockout HEK-293T cell lysate

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PARP1 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 2.
2 Images
Western blot - Human PARP1 knockout HEK-293T cell lysate (AB257017)
  • WB

Lab

Western blot - Human PARP1 knockout HEK-293T cell lysate (AB257017)

Lane 1 : Wild-type HEK-293T cell lysate (20µg)

Lane 2 : PARP1 knockout HEK-293T cell lysate (20µg)

Lanes 1- 2 : Merged signal (red and green). Green - ab32138 observed at 113 kDa. Red - loading control ab8245 observed at 37 kDa.

ab32138 Anti-PARP1 antibody [E102] was shown to specifically react with PARP1 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266598 (knockout cell lysate ab257017) was used. Wild-type and PARP1 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. ab32138 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 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-PARP1 antibody [E102] (<a href='/en-us/products/primary-antibodies/parp1-antibody-e102-ab32138'>ab32138</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

PARP1 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human PARP1 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-parp1-knockout-hek-293t-cell-line-ab266598'>ab266598</a>)

Predicted band size: 113 kDa

Observed band size: 113 kDa

false

Sanger Sequencing - Human PARP1 knockout HEK-293T cell lysate (AB257017)
  • Sanger seq

Unknown

Sanger Sequencing - Human PARP1 knockout HEK-293T cell lysate (AB257017)

Homozygous : 1 bp insertion in exon 2

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2.

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Complementary Products

Primary Antibodies

AB32064

Anti-Cleaved PARP1 antibody [E51]

RabMAb|Recombinant|KO Validated|Lab Essentials|20ul selling size

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cleaved PARP1 antibody [E51] (AB32064)

  • 4
  • 18
Proteins & Peptides

AB52101

Recombinant human Cleaved Caspase-3 protein (Active)

Western blot - Recombinant human Cleaved Caspase-3 protein (Active) (AB52101)

  • 0
  • 0
Proteins & Peptides

AB198766

Recombinant human PARP2 protein

Functional Studies - Recombinant human PARP2 protein (AB198766)

  • 0
  • 0
Proteins & Peptides

AB79663

Recombinant human PARP1 protein

Functional Studies - Recombinant human PARP1 protein (AB79663)

  • 0
  • 0
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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
PARP1
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.

PARP1 also known as poly(ADP-ribose) polymerase 1 is an enzyme that plays an important role in DNA repair processes. It detects DNA single-strand breaks and uses NAD+ as a substrate to add ADP-ribose polymers to itself and other proteins. This post-translational modification signals DNA repair machinery to the site of damage. PARP1 has a molecular weight of approximately 116 kDa. It is widely expressed in the nucleus of eukaryotic cells. PARP1 is often studied by western blotting techniques to analyze its expression and activation levels.
Biological function summary

Poly(ADP-ribose) polymerase 1 functions to maintain genomic stability by acting within the base excision repair complex. This complex is important for the detection and repair of DNA damage preventing the accumulation of mutations. By acting at sites of DNA stress PARP1 facilitates the binding of DNA repair proteins stabilizing the DNA structure during the repair process. This role is significant for cells that undergo frequent DNA replication or are exposed to high levels of genotoxic stress.

Pathways

The PARP1 protein is integral to the DNA damage response and repair pathway. It interacts with other proteins such as XRCC1 to coordinate repair activities at damaged DNA sites. Another important pathway involving PARP1 is the apoptosis pathway where excessive activation of PARP1 can lead to cell death due to depletion of cellular NAD+ and ATP. This indicates its dual role in both promoting cell survival through DNA repair and contributing to cell death when damage is irreparable.

Poly(ADP-ribose) polymerase 1 is strongly linked to cancer and neurodegenerative diseases. Its activity is heightened in many cancer types where cancer cells exploit PARP1 for survival by repairing DNA damage that would otherwise be lethal. Inhibitors of PARP1 are being developed as cancer therapies to target these survival mechanisms. Moreover overactivation of PARP1 in neurodegenerative disorders like Alzheimer's disease can lead to excessive energy consumption promoting neuronal cell damage. In these contexts PARP1 connects with proteins like BRCA1 in cancer or AIF in neurodegeneration illustrating its role in disease mechanisms.
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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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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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