Human MCL1 knockout HEK-293T cell line
Human MCL1 knockout HEK-293T cell line
- Advanced Validation
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MCL1 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 1 and 7 bp insertion in exon 1. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
別名を表示する
BCL2 related, Bcl-2-like protein 3, Bcl-2-related protein EAT/mcl1, Bcl2-L-3, EAT, Induced myeloid leukemia cell differentiation protein Mcl-1, MCL1-ES, MCL1L, MCL1S, MCL1_HUMAN, MGC104264, MGC1839, Myeloid Cell Leukemia 1, Myeloid cell leukemia ES, Myeloid cell leukemia sequence 1, Myeloid cell leukemia sequence 1 BCL2 related, Myeloid cell leukemia sequence 1 isoform 1, OTTHUMP00000032794, OTTHUMP00000032795, TM, mcl1/EAT
- WB
Lab
Western blot - Human MCL1 knockout HEK-293T cell line (AB266838)
Lanes 1-3 : Merged signal (red and green). Green - ab32087 observed at 37 kDa. Red - loading control ab7291 observed at 50 kDa.
ab32087 Anti-MCL1 antibody [Y37] was shown to specifically react with MCL1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266838 (knockout cell lysate ab256986) was used. Wild-type and MCL1 knockout samples were subjected to SDS-PAGE. ab32087 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4° 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-MCL1 antibody [Y37] (<a href='/products/primary-antibodies/mcl1-antibody-y37-ab32087'>ab32087</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK293T cell lysate at 20 µg
Lane 2:
MCL1 knockout HEK293T cell lysate at 20 µg
Lane 2:
Western blot - Human MCL1 knockout HEK-293T cell line (ab266838)
Lane 3:
Ramos cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution
Predicted band size: 37 kDa
Observed band size: 37 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human MCL1 knockout HEK-293T cell line (AB266838)
Allele-1 : 1 bp deletion in exon 1
- Sanger seq
Unknown
Sanger Sequencing - Human MCL1 knockout HEK-293T cell line (AB266838)
Allele-2 : 7 bp insertion in exon 1.
Reactivity data
製品の詳細
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.
製品内容
出荷温度及び保存条件
遺伝子名
遺伝子編集のタイプ
遺伝子編集の方法
ノックアウト検証方法
出荷温度
短期保存温度
長期保存温度
取り扱い方法
初回取り扱いガイドライン
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.
継代培養ガイドライン
- 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.
培養培地
DMEM (High Glucose) + 10% FBS
凍結保存培地
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
補足情報
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
MCL1 protein functions to maintain cell viability by counteracting pro-apoptotic stimuli. It interacts within the cell as part of a complex network involving other BCL-2 family proteins. MCL1 binds and sequesters BH3-only proteins and Bax therefore preventing them from initiating the apoptotic cascade. This interaction helps cells survive under stress conditions by temporarily inhibiting apoptosis.
Pathways
MCL1 plays a significant role in the intrinsic (mitochondrial) apoptosis pathway and the cell signaling pathways that facilitate cell survival and proliferation. In the mitochondrial pathway it works closely with other BCL-2 family members like BCL-xL and BCL-2 itself to control the permeability of the mitochondrial outer membrane and thereby influence cytochrome c release. The PI3K/AKT pathway can also regulate MCL1 leading to increases in its expression and stability which promotes cell survival.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
ターゲットの情報
Abcam 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