人MTAP knockout HeLa cell line
Human MTAP knockout HeLa cell line
- Advanced Validation
- 了解详情
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MTAP 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 1 and Insertion of the selection cassette in exon 1. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
查看别名
5' methylthioadenosine phosphorylase, 5''-methylthioadenosine phosphorylase, BDMF, DMSFH, DMSMFH, Epididymis luminal protein 249, HEL 249, LGMBF, MSAP, MTA phosphorylase, MTAP_HUMAN, MTAPase, MeSAdo phosphorylase, Methylthioadenosine phosphorylase, S methyl 5 thioadenosine phosphorylase, S methyl 5' thioadenosine phosphorylase, S-methyl-5''-thioadenosine phosphorylase, c86fus
- WB
Lab
Western blot - Human MTAP knockout HeLa cell line (AB265272)
Lanes 1-4 : Merged signal (red and green). Green - ab254265 observed at 32 kDa. Red - loading control, ab8245 observed at 37 kDa.
ab254265 Anti-MTAP antibody [EPR22570-76] was shown to specifically react with MTAP in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265272 (knockout cell lysate ab257194) was used. Wild-type and MTAP knockout samples were subjected to SDS-PAGE. ab254265 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 10000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-MTAP antibody [EPR22570-76] (<a href='/products/primary-antibodies/mtap-antibody-epr22570-76-ab254265'>ab254265</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
MTAP knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human MTAP knockout HeLa cell line (ab265272)
Lane 3:
HT-29 cell lysate at 20 µg
Lane 4:
A549 cell lysate at 20 µg
Predicted band size: 31 kDa
Observed band size: 32 kDa
false
- WB
Lab
Western blot - Human MTAP knockout HeLa cell line (AB265272)
anes 1- 4 : Merged signal (red and green). Green - ab126770 observed at 32 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab126770 Anti-MTAP antibody [EPR6893] was shown to specifically react with MTAP in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265272 (knockout cell lysate ab257194) was used. Wild-type and MTAP knockout samples were subjected to SDS-PAGE. ab126770 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 10000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-MTAP antibody [EPR6893] (<a href='/products/primary-antibodies/mtap-antibody-epr6893-ab126770'>ab126770</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
MTAP knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human MTAP knockout HeLa cell line (ab265272)
Lane 3:
HT-29 cell lysate at 20 µg
Lane 4:
A549 cell lysate at 20 µg
Predicted band size: 31 kDa
Observed band size: 32 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human MTAP knockout HeLa cell line (AB265272)
Allele-2 : Insertion of the selection cassette in exon 1.
- Sanger seq
Unknown
Sanger Sequencing - Human MTAP knockout HeLa cell line (AB265272)
Allele-1 : 1 bp insertion in exon 1.
- Sanger seq
Unknown
Sanger Sequencing - Human MTAP knockout HeLa cell line (AB265272)
Allele-3 : Insertion of the selection cassette in exon 1.
- Cell Culture
Unknown
Cell Culture - Human MTAP knockout HeLa cell line (AB265272)
Representative images of MTAP knockout HeLa cells, low and high confluency examples (top left and right respectively) and wild-type HeLa cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS XL Core microscope.
反应性数据
产品详情
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.
规格
性能和储存信息
基因名称
基因编辑类型
基因编辑方法
敲除验证
运输条件
推荐的短期储存条件
推荐的长期储存条件
处理步骤
初始处理指南
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.
传代培养指南
- 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.
培养基
DMEM (High Glucose) + 10% FBS
低温储藏试剂
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
补充信息
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
Methylthioadenosine phosphorylase plays a significant role in the salvage pathways for methionine and adenine critical for cellular growth and proliferation. MTAP operates as a part of a complex metabolic network involved in polyamine metabolism. In addition to its metabolic functions MTAP contributes to the regulation of the immune response and cell cycle. MTAP immunohistochemistry is often used to study its expression patterns in various tissues.
Pathways
Methylthioadenosine phosphorylase participates importantly in the polyamine biosynthesis and methionine salvage pathways. These pathways are integral for maintaining cellular homeostasis and nucleotide pools. MTAP works closely with proteins such as methionine adenosyltransferase (MAT) and adenosylmethionine decarboxylase (AMD). In concert they facilitate the regeneration of methionine highlighting MTAP's role in cellular adaptation to metabolic demands.
质量控制
STR 分析
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
细胞培养
生物安全等级
EU: 2 US: 2
贴壁/悬浮
Adherent
性别
Female
搭配使用产品
Primary Antibodies
AB126770
RabMAb|Recombinant|KO Validated|20ul selling size
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTAP antibody [EPR6893] (AB126770)](https://content.abcam.com/products/images/mtap-antibody-epr6893-ab126770--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img128866.gif)
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Abcam Product Promise
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