Human SETD2 (KMT3A / HYPB / HIF-1) knockout HeLa cell line
- Advanced Validation
- What is this?
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SETD2 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 insertion in exon 3 and 2 bp deletion in exon 3. 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
EC 2.1.1.43, FLJ16420, FLJ22472, FLJ23184, FLJ45883, HBP231, HIF-1, HSPC069, HYPB, Histone-lysine N-methyltransferase SETD2, Huntingtin interacting protein, Huntingtin interacting protein HYPB, Huntingtin yeast partner B, Huntingtin-binding protein, 231-KD, Huntingtin-interacting protein 1, Huntingtin-interacting protein B, KIAA1732, KMT3A, Lysine N-methyltransferase 3A, SET domain containing 2, SET domain-containing protein 2, SET2, SETD2_HUMAN, hSET2, p231HBP
- WB
Lab
Western blot - Human SETD2 (KMT3A / HYPB / HIF-1) knockout HeLa cell line (AB261792)
False colour image of Western blot : Anti-KMT3A / HYPB / HIF-1 antibody [EPR20927-67] staining at 1/1000 dilution shown in black; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution shown in red. In Western blot ab239350 was shown to bind specifically to KMT3A / HYPB / HIF-1. A band was observed at 120-280 440 kDa in wild-type HeLa cell lysates with no signal observed at this size in SETD2 knockout cell line ab261792 (knockout cell lysate ab257273). To generate this image wild-type and SETD2 knockout HeLa cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°. Blots were washed four times in TBS-T incubated with secondary antibodies for 1 h at room temperature washed again four times before development with Optiblot (ECL reagent ab133456) and imaged with 20 minutes exposure time. Secondary antibodies used were HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-KMT3A / HYPB / HIF-1 antibody [EPR20927-67] (<a href='/en-us/products/primary-antibodies/kmt3a-hypb-hif-1-antibody-epr20927-67-ab239350'>ab239350</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
SETD2 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human SETD2 (KMT3A / HYPB / HIF-1) knockout HeLa cell line (ab261792)
Lane 3:
HepG2 cell lysate at 20 µg
Lane 4:
Human Heart cell lysate at 20 µg
Predicted band size: 288 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human SETD2 (KMT3A / HYPB / HIF-1) knockout HeLa cell line (AB261792)
Allele-2 : 1 bp insertion in exon 3.
- Sanger seq
Unknown
Sanger Sequencing - Human SETD2 (KMT3A / HYPB / HIF-1) knockout HeLa cell line (AB261792)
Allele-1 : 2 bp deletion in exon 3.
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
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.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
KMT3A modifies chromatin structure to regulate gene expression. Its histone methylation function plays a critical role in the modulation of transcriptional activity ensuring that genes are turned on or off when necessary for cellular function. Additionally KMT3A forms part of complex protein networks that mediate gene expression changes in response to cellular signals. The precise regulation of this activity is vital for maintaining normal cellular function and responding to environmental cues.
Pathways
KMT3A engages in significant pathways such as the transcriptional regulation and epigenetic modification pathways. It interacts with various proteins including CoREST and LSD1 within these pathways to modulate gene expression and influence cellular differentiation. Through these interactions KMT3A ensures the correct genetic programs are executed aligning with developmental and homeostatic requirements.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
Primary Antibodies
AB239350
Anti-KMT3A / HYPB / HIF-1 antibody [EPR20927-67]
RabMAb|Recombinant|KO Validated|Trial Size
![Western blot - Anti-KMT3A / HYPB / HIF-1 antibody [EPR20927-67] (AB239350)](https://content.abcam.com/products/images/kmt3a-hypb-hif-1-antibody-epr20927-67-ab239350--western-blot-img167197.jpg)
- 0
- 0
Cell Lines & Lysates
AB255394
Human HIF1A (HIF-1 alpha) knockout HCT116 cell line
Advanced Validation
- 5
- 2
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