Human PSMB10 (MECL1) knockout HeLa cell line
- Advanced Validation
- What is this?
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PSMB10 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 1.
View Alternative Names
FLJ00366, LMP10, Low molecular mass protein 10, MECL1, MGC1665, Macropain subunit MECl-1, Multicatalytic endopeptidase complex subunit MECl-1, OTTHUMP00000174858, PSB10_HUMAN, PSMB 10, Proteasome (prosome macropain) subunit beta type 10, Proteasome MECl-1, Proteasome beta 10 subunit, Proteasome catalytic subunit 2i, Proteasome subunit MECL1, Proteasome subunit beta 7i, Proteasome subunit beta type-10, Proteasome subunit beta-2i, beta2i
- WB
Lab
Western blot - Human PSMB10 (MECL1) knockout HeLa cell line (AB264923)
Lanes 1-4 : Merged signal (red and green). Green - ab183506 observed at 29 kDa. Red - loading control ab8245 observed at 36 kDa.
ab183506 Anti-PSMB10/MECL1 antibody [EPR14902] was shown to specifically react with PSMB10/MECL1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264923 (knockout cell lysate ab258145) was used. Wild-type and PSMB10/MECL1 knockout samples were subjected to SDS-PAGE. ab183506 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) 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-PSMB10/MECL1 antibody [EPR14902] (<a href='/en-us/products/primary-antibodies/psmb10-mecl1-antibody-epr14902-ab183506'>ab183506</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
PSMB10 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human PSMB10 (MECL1) knockout HeLa cell line (ab264923)
Lane 3:
Daudi cell lysate at 20 µg
Lane 4:
Raji 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 1/10000 dilution
Predicted band size: 29 kDa
Observed band size: 29 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human PSMB10 (MECL1) knockout HeLa cell line (AB264923)
Homozygous : 1 bp deletion in exon 1.
Reactivity data
Product details
Recommended control: Human wild-type HeLa cell line (ab255448). 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 and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant 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
PSMB10/MECL1 plays a significant role in immune responses. It forms part of the immunoproteasome a variant of the proteasome complex which enhances the generation of antigenic peptides presented on MHC class I molecules. This function is essential for effective immune surveillance and initializing adaptive immunity. In particular PSMB10's influence on antigen processing positions it as an important player in modulating immune system activity.
Pathways
PSMB10/MECL1 participates in pathways associated with protein catabolism and immune signaling. It is involved in the ubiquitin-proteasome pathway which is critical for maintaining cellular protein balance. The function of PSMB10 within the immunoproteasome links it to immune signaling pathways. It has associations with other 20S proteasome subunits such as PSMB8 and PSMB9 which also contribute to the formation of the immunoproteasome complex.
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
Primary Antibodies
AB183506
Anti-PSMB10/MECL1 antibody [EPR14902]
RabMAb|Recombinant|KO Validated
![Western blot - Anti-PSMB10/MECL1 antibody [EPR14902] (AB183506)](https://content.abcam.com/products/images/psmb10-mecl1-antibody-epr14902-ab183506--western-blot-img5033.jpg)
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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