Human B2M (beta 2 Microglobulin) knockout Hep G2 cell line
- Advanced Validation
- What is this?
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(0 Publication)
- WB
Lab
Western blot - Human B2M (beta 2 Microglobulin) knockout Hep G2 cell line (AB262325)
Lanes 1-4 : Merged signal (red and green). Green - ab218230 observed at 14 kDa. Red - loading control ab8245 observed at 36 kDa.
ab218230 Anti-beta 2 Microglobulin antibody [EPR21752-214] was shown to specifically react with beta 2 Microglobulin in wild-type HepG2 cells. Loss of signal was observed when knockout cell line ab262325 (knockout cell lysate ab256846) was used. Wild-type and beta 2 Microglobulin knockout samples were subjected to SDS-PAGE. ab218230 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at 1 in 500 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-beta 2 Microglobulin antibody [EPR21752-214] (<a href='/en-us/products/primary-antibodies/beta-2-microglobulin-antibody-epr21752-214-ab218230'>ab218230</a>) at 1/500 dilution
Lane 1:
Wild-type HepG2 cell lysate at 20 µg
Lane 2:
B2M knockout HepG2 cell lysate at 20 µg
Lane 2:
Western blot - Human B2M (beta 2 Microglobulin) knockout Hep G2 cell line (ab262325)
Lane 3:
HeLa cell lysate at 20 µg
Lane 4:
Jurkat 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: 14 kDa
Observed band size: 14 kDa
false
- WB
Lab
Western blot - Human B2M (beta 2 Microglobulin) knockout Hep G2 cell line (AB262325)
Lanes 1-4 : Merged signal (red and green). Green - ab75853 observed at 14 kDa. Red - loading control ab8245 observed at 36 kDa.
ab75853 Anti-beta 2 Microglobulin antibody [EP2978Y] was shown to specifically react with beta 2 Microglobulin in wild-type HepG2 cells. Loss of signal was observed when knockout cell line ab262325 (knockout cell lysate ab256846) was used. Wild-type and beta 2 Microglobulin knockout samples were subjected to SDS-PAGE. ab75853 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-beta 2 Microglobulin antibody [EP2978Y] (<a href='/en-us/products/primary-antibodies/beta-2-microglobulin-antibody-ep2978y-ab75853'>ab75853</a>) at 1/1000 dilution
Lane 1:
Wild-type HepG2 cell lysate at 20 µg
Lane 2:
B2M knockout HepG2 cell lysate at 20 µg
Lane 2:
Western blot - Human B2M (beta 2 Microglobulin) knockout Hep G2 cell line (ab262325)
Lane 3:
HeLa cell lysate at 20 µg
Lane 4:
Jurkat 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: 14 kDa
Observed band size: 14 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human B2M (beta 2 Microglobulin) knockout Hep G2 cell line (AB262325)
Allele-1 : 1 bp insertion in exon1
- Sanger seq
Unknown
Sanger Sequencing - Human B2M (beta 2 Microglobulin) knockout Hep G2 cell line (AB262325)
Allele-2 : Insertion of the selection cassette in exon 1.
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Advanced Validation
- 0
- 0
Reactivity data
Product details
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
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
MEM + 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
Beta-2-microglobulin is important for the stability and transport of MHC class I molecules to the cell surface. As part of the MHC class I complex B2M assists in binding peptides allowing immune cells to identify and target pathogen-infected cells. Without B2M the MHC class I molecules are not properly expressed on the cell surface disrupting immune recognition. In laboratory settings researchers often use anti-beta-2-microglobulin antibodies to investigate its role in MHC class I function.
Pathways
Beta-2-microglobulin interacts significantly with the immune system most notably in the antigen processing and presentation pathway. It works alongside proteins such as the heavy chain of MHC class I. B2M is important in the pathway that involves the transport of antigens to the endoplasmic reticulum where they are loaded onto MHC class I molecules for inspection by cytotoxic T cells. Another related pathway is the tapasin-mediated processing of antigen peptides highlighting the indispensable role of B2M in immune response regulation.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Target data
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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