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AB265770

Human SETD7 (SET7) knockout HeLa cell line

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SETD7 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 2 and 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'.
4 Images
Western blot - Human SETD7 (SET7) knockout HeLa cell line (AB265770)
  • WB

Unknown

Western blot - Human SETD7 (SET7) knockout HeLa cell line (AB265770)

Blocking and dilution buffer : Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBST

Lanes 1-3 : Merged signal (red and green). Green - ab274416 observed at 50 kDa. Red - loading control ab8245 observed at 36 kDa.

ab274416 Anti-SET7 antibody [EPR23775-142] was shown to specifically react with SET7 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265770 (knockout cell lysate ab257668) was used. Wild-type and SET7 knockout samples were subjected to SDS-PAGE. ab274416 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours 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-SET7 antibody [EPR23775-142] (<a href='/en-us/products/primary-antibodies/set7-antibody-epr23775-142-ab274416'>ab274416</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

SET7 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human SETD7 (SET7) knockout HeLa cell line (ab265770)

Lane 3:

Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/10000 dilution

Predicted band size: 41 kDa

Observed band size: 50 kDa

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Western blot - Human SETD7 (SET7) knockout HeLa cell line (AB265770)
  • WB

Lab

Western blot - Human SETD7 (SET7) knockout HeLa cell line (AB265770)

Lanes 1-3 : Merged signal (red and green). Green - ab124708 observed at 50 kDa. Red - loading control ab8245 observed at 36 kDa.

ab124708 Anti-SET7 antibody [EPR5574] was shown to specifically react with SET7 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265770 (knockout cell lysate ab257668) was used. Wild-type and SET7 knockout samples were subjected to SDS-PAGE. ab124708 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours 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-SET7 antibody [EPR5574] (<a href='/en-us/products/primary-antibodies/set7-antibody-epr5574-ab124708'>ab124708</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

SETD7 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human SETD7 (SET7) knockout HeLa cell line (ab265770)

Lane 3:

U-2 OS cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 41 kDa

Observed band size: 50 kDa

false

Sanger Sequencing - Human SETD7 (SET7) knockout HeLa cell line (AB265770)
  • Sanger seq

Unknown

Sanger Sequencing - Human SETD7 (SET7) knockout HeLa cell line (AB265770)

Allele-1 : 1 bp deletion in exon 2.

Sanger Sequencing - Human SETD7 (SET7) knockout HeLa cell line (AB265770)
  • Sanger seq

Unknown

Sanger Sequencing - Human SETD7 (SET7) knockout HeLa cell line (AB265770)

Allele-2 : 1 bp insertion in exon 2.

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Form

Liquid

form

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 2 and 1 bp insertion in exon 2

Antibiotic resistance

Puromycin 1µg/mL

Disease

Adenocarcinoma

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Complementary Products

Primary Antibodies

AB124708

Anti-SET7 antibody [EPR5574]

RabMAb|Recombinant|KO Validated

Immunoprecipitation - Anti-SET7 antibody [EPR5574] (AB124708)

  • 0
  • 0
Primary Antibodies

AB219857

Anti-FRMD8 antibody

Immunocytochemistry/ Immunofluorescence - Anti-FRMD8 antibody (AB219857)

  • 0
  • 0
Primary Antibodies

AB272656

Anti-COBLL1 antibody

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COBLL1 antibody (AB272656)

  • 0
  • 0
Primary Antibodies

AB235083

Anti-ALG9 antibody

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ALG9 antibody (AB235083)

  • 0
  • 0
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Reactivity data

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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.

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What's included?

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Properties and storage information

Gene name
SETD7
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C
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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.

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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

SET7 also known as SETD7 or KMT7 is a lysine methyltransferase that adds methyl groups to lysine residues on target proteins influencing their function and stability. This protein has a molecular mass of approximately 55 kDa. SET7 is expressed in a variety of tissues including the heart liver and pancreas suggesting its versatile role across different biological systems.
Biological function summary

SET7 contributes to gene regulation by methylating histone H3 at lysine 4 (H3K4) a mark generally associated with transcriptional activation. It also methylates non-histone proteins such as p53 TAF10 and STAT3 affecting their activity and interactions. SET7 operates distinctively and does not need to be part of any complex to fulfill its function effectively in diverse cellular processes.

Pathways

SET7 modulates transcriptional pathways such as the p53 signaling pathway and the NF-kB pathway. It interacts with the p53 protein affecting cell cycle regulation and the response to DNA damage. The modification of NF-kB by SET7 can influence inflammation and immune responses. Through these pathways SET7 plays a role in maintaining cellular homeostasis and response to stress.

SET7 is involved in conditions like cancer and cardiovascular diseases. Its methylation of p53 links it to tumor suppression activities while its impact on NF-kB relates to inflammatory responses. Dysregulation of SET7 activity can lead to abnormal cell proliferation and inflammation pointing to its potential as a target for therapeutic intervention.
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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'.

For licensing inquiries, please contact partnerships@abcam.com