Human SP1 knockout HeLa cell line
- Advanced Validation
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- ChIP-seq
Supplier Data
ChIP-sequencing - Human SP1 knockout HeLa cell line (AB265519)
Chromatin was prepared from Human SP1 knockout HeLa cell line (ab265519) cells overexpressing either mNeonGreen or mNeonGreen-SP1. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 107 cells and 8 μg ab321887 [EPR28835-76]. Assay quality control was conducted using 8 μg anti-SP1 (ab231778) on the same cell lines. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.
- ChIP-seq
Supplier Data
ChIP-sequencing - Human SP1 knockout HeLa cell line (AB265519)
Chromatin was prepared from Human SP1 knockout HeLa cell line (ab265519) cells overexpressing either mNeonGreen or mNeonGreen-SP1. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 107 cells and 8 μg ab321887 [EPR28835-76]. Assay quality control was conducted using 8 μg anti-SP1 (ab231778) on the same cell lines. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.
- ChIP-seq
Supplier Data
ChIP-sequencing - Human SP1 knockout HeLa cell line (AB265519)
Chromatin was prepared from Human SP1 knockout HeLa cell line (ab265519) cells overexpressing either mNeonGreen or mNeonGreen-SP1. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 107 cells and 8 μg ab321887 [EPR28835-76]. Assay quality control was conducted using 8 μg anti-SP1 (ab231778) on the same cell lines. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.
- WB
Lab
Western blot - Human SP1 knockout HeLa cell line (AB265519)
Lanes 1-2 : Merged signal (red and green). Green - ab231778 observed at 100 kDa. Red - loading control ab8245 observed at 37 kDa.
ab231778 Anti-SP1 antibody [EPR22648-50] was shown to specifically react with SP1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265519 (knockout cell lysate ab257698) was used. Wild-type and SP1 knockout samples were subjected to SDS-PAGE. ab231778 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 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-SP1 antibody [EPR22648-50] - ChIP Grade (<a href='/en-us/products/primary-antibodies/sp1-antibody-epr22648-50-chip-grade-ab231778'>ab231778</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
SP1 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human SP1 knockout HeLa cell line (ab265519)
Predicted band size: 81 kDa
Observed band size: 100 kDa
false
- WB
Lab
Western blot - Human SP1 knockout HeLa cell line (AB265519)
Lanes 1-2 : Merged signal (red and green). Green - ab124804 observed at 100 kDa. Red - loading control ab8245 observed at 37 kDa.
ab124804 Anti-SP1 antibody [EPR6662(B)] was shown to specifically react with SP1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265519 (knockout cell lysate ab257698) was used. Wild-type and SP1 knockout samples were subjected to SDS-PAGE. ab124804 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 5000 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-SP1 antibody [EPR6662(B)] (<a href='/en-us/products/primary-antibodies/sp1-antibody-epr6662b-ab124804'>ab124804</a>) at 1/5000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
SP1 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human SP1 knockout HeLa cell line (ab265519)
Predicted band size: 81 kDa
Observed band size: 100 kDa
false
- ChIC/CUT&RUN-seq
Supplier Data
ChIC/CUT&RUN sequencing - Human SP1 knockout HeLa cell line (AB265519)
ChIC/CUT&RUN was performed using pAG-MNase at a final concentration of 700 ng/mL. 2.5 x 105 Human SP1 knockout HeLa cells (ab265519) over expressing either mNeonGreen or mNeonGreen-SP1 were used along with 5 μg of ab321887[EPR28835-76]. Assay quality control was conducted using 5 μg anti-SP1(ab231778) on the same cell lines. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.
The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
- ChIC/CUT&RUN-seq
Supplier Data
ChIC/CUT&RUN sequencing - Human SP1 knockout HeLa cell line (AB265519)
ChIC/CUT&RUN was performed using pAG-MNase at a final concentration of 700 ng/mL. 2.5 x 105 Human SP1 knockout HeLa cells (ab265519) over expressing either mNeonGreen or mNeonGreen-SP1 were used along with 5 μg of ab321887[EPR28835-76]. Assay quality control was conducted using 5 μg anti-SP1(ab231778) on the same cell lines. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.
The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
- ChIC/CUT&RUN-seq
Supplier Data
ChIC/CUT&RUN sequencing - Human SP1 knockout HeLa cell line (AB265519)
ChIC/CUT&RUN was performed using pAG-MNase at a final concentration of 700 ng/mL. 2.5 x 105 Human SP1 knockout HeLa cells (ab265519) over expressing either mNeonGreen or mNeonGreen-SP1 were used along with 5 μg of ab321887[EPR28835-76]. Assay quality control was conducted using 5 μg anti-SP1(ab231778) on the same cell lines. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.
The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
- Sanger seq
Unknown
Sanger Sequencing - Human SP1 knockout HeLa cell line (AB265519)
Homozygous : Insertion of the selection cassette in exon 2.
Complementary Products
Primary Antibodies
AB231778
Anti-SP1 antibody [EPR22648-50] - ChIP Grade
BOND RX™ Validated|Recombinant|KO Validated|Advanced Validation|RabMAb
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SP1 antibody [EPR22648-50] - ChIP Grade (AB231778)](https://content.abcam.com/products/images/sp1-antibody-epr22648-50-chip-grade-ab231778--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img67547.jpg)
- 0
- 0
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
Handling procedures
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.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
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.
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.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
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.
Quality control
STR analysis
D16S539, TPOX, CSF1PO, D13S317, D7S820, D5S818, TH01
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
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