人BCKDHA knockout HEK-293T cell line
Human BCKDHA knockout HEK-293T cell line
- Advanced Validation
- 了解详情
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BCKDHA KO cell line available to order. KO validated by. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 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
2 oxoisovalerate dehydrogenase (lipoamide), 2-oxoisovalerate dehydrogenase subunit alpha, mitochondrial, BCKDE1A, BCKDH E1-alpha, Branched chain alpha keto acid dehydrogenase E1 component alpha polypeptide, Branched chain keto acid dehydrogenase E1 alpha polypeptide, Branched chain keto acid dehydrogenase E1, alpha polypeptide (maple syrup urine disease), Branched-chain alpha-keto acid dehydrogenase E1 component alpha chain, FLJ45695, MSU, MSUD1, ODBA_HUMAN, OVD1A
- WB
Lab
Western blot - Human BCKDHA knockout HEK-293T cell line (AB266439)
False colour image of Western blot : Anti-BCKDHA antibody staining at 1/500 dilution shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution shown in red. In Western blot ab126173 was shown to bind specifically to BCKDHA. A band was observed at 48 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in BCKDHA knockout cell line ab266439 (knockout cell lysate ab258324). To generate this image wild-type and BCKDHA knockout HEK-293T 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 then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-BCKDHA antibody (<a href='/products/primary-antibodies/bckdha-antibody-ab126173'>ab126173</a>) at 1/500 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
BCKDHA knockout HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human BCKDHA knockout HEK-293T cell line (ab266439)
Lane 3:
Human Heart cell lysate at 20 µg
Lane 4:
Daudi cell lysate at 20 µg
Predicted band size: 50 kDa
Observed band size: 48 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human BCKDHA knockout HEK-293T cell line (AB266439)
Homozygous : Insertion of the selection cassette in exon1
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
合子性
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.
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
AB305168
Anti-BCKDHA antibody [EPR27003-11]
BOND RX™ Validated|20ul selling size|RabMAb|Recombinant|KO Validated
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BCKDHA antibody [EPR27003-11] (AB305168)](https://content.abcam.com/products/images/bckdha-antibody-epr27003-11-ab305168--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img405421.jpg)
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Abcam Product Promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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