Human SERPINB5 (MASPIN) knockout HeLa cell line
- Advanced Validation
- What is this?
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SERPINB5 KO cell line available to order. KO validated by. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2.
View Alternative Names
PI-5, Peptidase inhibitor 5, Protease inhibitor 5, SPB5_HUMAN, Serine (or cysteine) peptidase inhibitor, clade B, member 5, Serpin B5, Serpin peptidase inhibitor, Serpin peptidase inhibitor, clade B (ovalbumin), member 5, Spi5, Spi7, protease inhibitor 5 (maspin), serine (or cysteine) proteinase inhibitor, clade B (ovalbumin), member 5, serpin family B member 5
- WB
Lab
Western blot - Human SERPINB5 (MASPIN) knockout HeLa cell line (AB264750)
Lanes 1 - 4 : Merged signal (red and green). Green - ab65136 observed at 42 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
ab65136 was shown to react with MASPIN in wild-type cells in Western blot with loss of signal observed in SERPINB5 knockout cell line ab264750 (SERPINB5 knockout cell lysate ab258659). Wild-type and SERPINB5 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with ab65136 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4° at a 1/500 dilution and a 1/20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1/20000 dilution for 1 h at room temperature before imaging.
All lanes:
Western blot - Anti-MASPIN antibody (<a href='/en-us/products/primary-antibodies/maspin-antibody-ab65136'>ab65136</a>) at 1/500 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
SERPINB5 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human SERPINB5 (MASPIN) knockout HeLa cell line (ab264750)
Lane 3:
HaCaT cell lysate at 20 µg
Lane 4:
Caco-2 cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution
Predicted band size: 42 kDa
Observed band size: 42 kDa
false
- WB
Lab
Western blot - Human SERPINB5 (MASPIN) knockout HeLa cell line (AB264750)
Lanes 1 - 4 : Merged signal (red and green). Green - ab272858 observed at 42 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
ab272858 was shown to react with MASPIN in wild-type cells in Western blot with loss of signal observed in SERPINB5 knockout cell line ab264750 (SERPINB5 knockout cell lysate ab258659). Wild-type and SERPINB5 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with ab272858 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4° at a 1/500 dilution and a 1/20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1/20000 dilution for 1 h at room temperature before imaging.
All lanes:
Western blot - Anti-MASPIN antibody (<a href='/en-us/products/primary-antibodies/maspin-antibody-ab272858'>ab272858</a>) at 1/500 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
SERPINB5 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human SERPINB5 (MASPIN) knockout HeLa cell line (ab264750)
Lane 3:
HaCaT cell lysate at 20 µg
Lane 4:
Caco-2 cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution
Predicted band size: 42 kDa
Observed band size: 42 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human SERPINB5 (MASPIN) knockout HeLa cell line (AB264750)
Homozygous : 1 bp insertion in exon 2.
Reactivity data
Product details
Recommended control: Human wild-type HeLa cell line (ab255928). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
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
Zygosity
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
MASPIN influences apoptosis and cell motility playing a critical role in tumor suppression. It does not form part of a complex but interacts directly with extracellular matrix components. MASPIN can inhibit angiogenesis by reducing the proliferation and migration of endothelial cells. Scientists have noted its role in maintaining normal cell architecture and inhibiting tumor cell invasion.
Pathways
Researchers place MASPIN in important signaling networks impacting cancer progression and metastasis. It participates in the PI3K/AKT pathway thereby influencing cell growth and survival. MASPIN also intersects with TGF-β signaling and related proteins like SMAD which mediate growth arrest and apoptosis. By altering these pathways MASPIN helps modulate responses to cellular stress and inhibit lethal growth in cancer cells.
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
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