Human PARP1 knockout HEK-293T cell line
- Advanced Validation
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PARP1 KO cell line available to order. KO validated by Flow Cytometry, Immunocytochemistry, Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
View Alternative Names
Poly [ADP-ribose] polymerase 1, PPOL, Poly[ADP-ribose] synthase 1, PARP1, Protein poly-ADP-ribosyltransferase PARP1, PARP-1, NAD(+) ADP-ribosyltransferase 1, ADPRT 1, ADP-ribosyltransferase diphtheria toxin-like 1, ADPRT, ARTD1, DNA ADP-ribosyltransferase PARP1
- Flow Cyt (Intra)
Supplier Data
Flow Cytometry (Intracellular) - Human PARP1 knockout HEK-293T cell line (AB266598)
HEK293T wild-type and PARP1 knockout cells (ab266598) were labelled with a green or violet, fluorescent dye, respectively. Wild-type and knockout cells were mixed in a 1 : 1 ratio, fixed in 4% PFA and permeabilized in 0.1% Triton X-100.
Antibody staining was quantified using the Attune NxT Flow Cytometer with representative images showing the staining intensity in the knockout population (pink histogram, dashed line) compared to the wild-type cells (green histogram, solid line). Histograms with dotted lines represent secondary antibody-only controls in both wild-type and knockout cells.
Antibody dilution : 0.5 μg/μL.
This data was provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
- Flow Cyt (Intra)
Supplier Data
Flow Cytometry (Intracellular) - Human PARP1 knockout HEK-293T cell line (AB266598)
HEK293T wild-type and PARP1 knockout cells (ab266598) were labelled with a green or violet, fluorescent dye, respectively. Wild-type and knockout cells were mixed in a 1 : 1 ratio, fixed in 4% PFA and permeabilized in 0.1% Triton X-100.
Antibody staining was quantified using the Attune NxT Flow Cytometer with representative images showing the staining intensity in the knockout population (pink histogram, dashed line) compared to the wild-type cells (green histogram, solid line). Histograms with dotted lines represent secondary antibody-only controls in both wild-type and knockout cells.
Antibody dilution : 0.1 μg/μL.
This data was provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
- ICC/IF
Supplier Data
Immunocytochemistry/ Immunofluorescence - Human PARP1 knockout HEK-293T cell line (AB266598)
ab32071 was shown to react with PARP1 in wild-type HEK293T cells in immunocytochemistry with loss of signal observed in PARP1 knockout cell line ab266598. Wild-type and knockout cells were mixed and pelleted at a 1 : 1 ratio on coverslips. The cells were fixed with 4% paraformaldehyde (15 min) then permeabilized with 0.1% Triton X-100 (10min) and then blocked with 1x PBS, 0.01% Triton X-100, 5% BSA, 5% NGS. The cells were then incubated with ab32071 at 1/50 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat anti-rabbit secondary antibody to (Alexa Fluor® 555) at 0.5 μg/ml. Acquisition of the green (wild-type), red (antibody staining) and far-red (knockout) channels was performed. Representative grayscale images of the red channel are shown. Wild-type and knockout cells are outlined with yellow and magenta dashed line, respectively. Schematic representation of the mosaic strategy used is shown on the bottom-right panel. Image was acquired with a Zeiss (LSM-880).
This data was provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
- ICC/IF
Supplier Data
Immunocytochemistry/ Immunofluorescence - Human PARP1 knockout HEK-293T cell line (AB266598)
ab32378 was shown to react with PARP1 in wild-type HEK293T cells in immunocytochemistry with loss of signal observed in PARP1 knockout cell line ab266598. Wild-type and knockout cells were mixed and pelleted at a 1 : 1 ratio on coverslips. The cells were fixed with 4% paraformaldehyde (15 min) then permeabilized with 0.1% Triton X-100 (10min) and then blocked with 1x PBS, 0.01% Triton X-100, 5% BSA, 5% NGS. The cells were then incubated with ab32378 at 1/500 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat anti-rabbit secondary antibody to (Alexa Fluor® 555) at 0.5 μg/ml. Acquisition of the green (wild-type), red (antibody staining) and far-red (knockout) channels was performed. Representative grayscale images of the red channel are shown. Wild-type and knockout cells are outlined with yellow and magenta dashed line, respectively. Schematic representation of the mosaic strategy used is shown on the bottom-right panel. Image was acquired with a Zeiss (LSM-880).
This data was provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
- ICC/IF
Supplier Data
Immunocytochemistry/ Immunofluorescence - Human PARP1 knockout HEK-293T cell line (AB266598)
ab32138 was shown to react with PARP1 in wild-type HEK293T cells in immunocytochemistry with loss of signal observed in PARP1 knockout cell line ab266598. Wild-type and knockout cells were mixed and pelleted at a 1 : 1 ratio on coverslips. The cells were fixed with 4% paraformaldehyde (15 min) then permeabilized with 0.1% Triton X-100 (10min) and then blocked with 1x PBS, 0.01% Triton X-100, 5% BSA, 5% NGS. The cells were then incubated with ab32138 at 1/50 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat anti-rabbit secondary antibody to (Alexa Fluor® 555) at 0.5 μg/ml. Acquisition of the green (wild-type), red (antibody staining) and far-red (knockout) channels was performed. Representative grayscale images of the red channel are shown. Wild-type and knockout cells are outlined with yellow and magenta dashed line, respectively. Schematic representation of the mosaic strategy used is shown on the bottom-right panel. Image was acquired with a Zeiss (LSM-880).
This data was provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
- Flow Cyt (Intra)
Supplier Data
Flow Cytometry (Intracellular) - Human PARP1 knockout HEK-293T cell line (AB266598)
HEK293T wild-type and PARP1 knockout cells (ab266598) were labelled with a green or violet, fluorescent dye, respectively. Wild-type and knockout cells were mixed in a 1 : 1 ratio, fixed in 4% PFA and permeabilized in 0.1% Triton X-100.
Antibody staining was quantified using the Attune NxT Flow Cytometer with representative images showing the staining intensity in the knockout population (pink histogram, dashed line) compared to the wild-type cells (green histogram, solid line). Histograms with dotted lines represent secondary antibody-only controls in both wild-type and knockout cells.
Antibody dilution : 0.1 μg/μL.
This data was provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
- WB
Lab
Western blot - Human PARP1 knockout HEK-293T cell line (AB266598)
Lanes 1- 2 : Merged signal (red and green). Green - ab32138 observed at 113 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab32138 was shown to 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 HEK-293T and PARP1 knockout HEK-293T cell lysates were subjected to SDS-PAGE. ab32138 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4° at a 1 in 1000 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-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 (ab266598)
Predicted band size: 113 kDa
Observed band size: 113 kDa
false
- WB
Supplier Data
Western blot - Human PARP1 knockout HEK-293T cell line (AB266598)
ab32378 was shown to react with PARP1 in wild-type HEK293T cells in Western blot with loss of signal observed in PARP1 knockout cell line ab266598. Wild-type HEK293T and PARP1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab32378 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.
This data was provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
All lanes:
Western blot - Anti-PARP1 antibody [Y17] (<a href='/en-us/products/primary-antibodies/parp1-antibody-y17-ab32378'>ab32378</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK293T lysate at 30 µg
Lane 2:
PARP1 knock-out HEK293T lysate at 30 µg
Lane 2:
Western blot - Human PARP1 knockout HEK-293T cell line (ab266598)
Predicted band size: 113 kDa
false
- WB
Supplier Data
Western blot - Human PARP1 knockout HEK-293T cell line (AB266598)
ab32071 was shown to react with PARP1 in wild-type HEK293T cells in Western blot with loss of signal observed in PARP1 knockout cell line ab266598. Wild-type HEK293T and PARP1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab32071 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.
This data was provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
All lanes:
Western blot - Anti-PARP1 antibody [E78] (<a href='/en-us/products/primary-antibodies/parp1-antibody-e78-ab32071'>ab32071</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK293T lysate at 30 µg
Lane 2:
PARP1 knock-out HEK293T lysate at 30 µg
Lane 2:
Western blot - Human PARP1 knockout HEK-293T cell line (ab266598)
Predicted band size: 113 kDa
false
- WB
Supplier Data
Western blot - Human PARP1 knockout HEK-293T cell line (AB266598)
ab32138 was shown to react with PARP1 in wild-type HEK293T cells in Western blot with loss of signal observed in PARP1 knockout cell line ab266598. Wild-type HEK293T and PARP1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab32138 overnight at 4 °C at a 1/5000 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.
This data was provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
All lanes:
Western blot - Anti-PARP1 antibody [E102] (<a href='/en-us/products/primary-antibodies/parp1-antibody-e102-ab32138'>ab32138</a>) at 1/5000 dilution
Lane 1:
Wild-type HEK293T lysate at 30 µg
Lane 2:
PARP1 knock-out HEK293T lysate at 30 µg
Lane 2:
Western blot - Human PARP1 knockout HEK-293T cell line (ab266598)
Predicted band size: 113 kDa
false
- Sanger seq
Lab
Sanger Sequencing - Human PARP1 knockout HEK-293T cell line (AB266598)
Sequencing chromatogram displaying sequence edit in exon 2
- Sanger seq
Unknown
Sanger Sequencing - Human PARP1 knockout HEK-293T cell line (AB266598)
Homozygous : 1 bp insertion in exon 2
- Cell Culture
Unknown
Cell Culture - Human PARP1 knockout HEK-293T cell line (AB266598)
Representative images of PARP1 knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using an EVOS M5000 microscope.
Complementary Products
Reactivity data
Product details
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Zygosity
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Initial handling guidelines
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 guidelines
- 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.
Culture medium
DMEM (High Glucose) + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Target data
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com