人GPI (AMF) knockout HEK-293T cell line
Human GPI (AMF) knockout HEK-293T cell line
- Advanced Validation
- 了解详情
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Human GPI (AMF) knockout HEK-293T cell line available to order. Recommended control: Human wild-type HEK293T cell line (ab255449).
查看别名
AMF, Aurocrine motility factor, Autocrine motility factor, DKFZp686C13233, EC 5.3.1.9, G6PI_HUMAN, GNPI, GPI, Glucose phosphate isomerase, Glucose-6-phosphate isomerase, Gpi1, Hexose monophosphate isomerase, Hexosephosphate isomerase, Neuroleukin, Oxoisomerase, PGI, PHI, Phosphoglucose isomerase, Phosphohexomutase, Phosphohexose isomerase, Phosphosaccharomutase, SA-36, Sperm antigen 36
- WB
Lab
Western blot - Human GPI (AMF) knockout HEK-293T cell line (AB266834)
Lanes 1-4 : Merged signal (red and green). Green - ab167394 observed at 63 kDa. Red - loading control ab8245 observed at 36 kDa.
ab167394 Anti-AMF antibody [EPR11663(B)] was shown to specifically react with AMF in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266834 (knockout cell lysate ab257458) was used. Wild-type and AMF knockout samples were subjected to SDS-PAGE. ab167394 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-AMF antibody [EPR11663(B)] (<a href='/products/primary-antibodies/amf-antibody-epr11663b-ab167394'>ab167394</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK293T cell lysate at 20 µg
Lane 2:
GPI knockout HEK293T cell lysate at 20 µg
Lane 2:
Western blot - Human GPI (AMF) knockout HEK-293T cell line (ab266834)
Lane 3:
HepG2 cell lysate at 20 µg
Lane 4:
HeLa 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: 63 kDa
Observed band size: 63 kDa
false
- WB
Lab
Western blot - Human GPI (AMF) knockout HEK-293T cell line (AB266834)
Lanes 1 - 2 : Merged signal (red and green). Green - ab66340 observed at 63 kDa. Red - loading control ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37 kDa.
ab66340 was shown to react with AMF in wild-type HEK-293T cells in Western blot with loss of signal observed in GPI knockout cell line ab266834 (GPI knockout cell lysate ab257458). Wild-type HEK-293T and GPI knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab66340 and ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4 ° at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
All lanes:
Western blot - Anti-AMF antibody [1B7D7] (<a href='/products/primary-antibodies/amf-antibody-1b7d7-ab66340'>ab66340</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
GPI knockout HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human GPI (AMF) knockout HEK-293T cell line (ab266834)
Predicted band size: 63 kDa
Observed band size: 63 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human GPI (AMF) knockout HEK-293T cell line (AB266834)
Homozygous : 76 bp deletion in exon2
反应性数据
产品详情
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
Abcam Product Promise
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