Human B2M (beta 2 Microglobulin) knockout HEK-293T cell line (ab266828)
Overview
-
Product name
Human B2M (beta 2 Microglobulin) knockout HEK-293T cell line
See all beta 2 Microglobulin lysates -
Parental Cell Line
HEK293T -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1 -
Passage number
<20 -
Knockout validation
Sanger Sequencing -
Tested applications
Suitable for: Sanger Sequencing, Cell Culture, WBmore details -
Biosafety level
2 -
General notes
Recommended control: Human wild-type HEK293T cell line (ab255449). 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.
Cryopreservation cell medium: Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Culture medium: DMEM (High Glucose) + 10% FBS
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. Based on cell count, seed cells in an appropriate cell culture 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.
4. Incubate the culture at 37°C incubator with 5% CO2. 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.
- A partial media change 24 hours prior to subculture may be helpful to encourage growth, if required.
- Cells should be passaged when they have achieved 80-90% confluence.
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.
Properties
-
Number of cells
1 x 106 cells/vial, 1 mL -
Viability
~80% -
Adherent /Suspension
Adherent -
Tissue
Kidney -
Cell type
epithelial -
STR Analysis
Amelogenin X D5S818: 8, 9 D13S317: 12, 14 D7S820: 11 D16S539: 9, 13 vWA: 16, 19 TH01: 7, 9.3 TPOX: 11 CSF1PO: 11, 12 -
Antibiotic resistance
Puromycin 1.00µg/ml -
Mycoplasma free
Yes -
Storage instructions
Shipped on Dry Ice. Store in liquid nitrogen. -
Storage buffer
Constituents: 8.7% Dimethylsulfoxide, 2% Cellulose, methyl ether -
Research areas
Target
-
Function
Component of the class I major histocompatibility complex (MHC). Involved in the presentation of peptide antigens to the immune system. -
Involvement in disease
Defects in B2M are the cause of hypercatabolic hypoproteinemia (HYCATHYP) [MIM:241600]. Affected individuals show marked reduction in serum concentrations of immunoglobulin and albumin, probably due to rapid degradation.
Note=Beta-2-microglobulin may adopt the fibrillar configuration of amyloid in certain pathologic states. The capacity to assemble into amyloid fibrils is concentration dependent. Persistently high beta(2)-microglobulin serum levels lead to amyloidosis in patients on long-term hemodialysis. -
Sequence similarities
Belongs to the beta-2-microglobulin family.
Contains 1 Ig-like C1-type (immunoglobulin-like) domain. -
Post-translational
modificationsGlycation of Ile-21 is observed in long-term hemodialysis patients. -
Cellular localization
Secreted. Detected in serum and urine. - Information by UniProt
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab266828 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| Sanger Sequencing |
Use at an assay dependent concentration.
|
|
| Cell Culture |
Use at an assay dependent concentration.
|
|
| WB |
Use at an assay dependent concentration.
|
| Notes |
|---|
|
Sanger Sequencing
Use at an assay dependent concentration. |
|
Cell Culture
Use at an assay dependent concentration. |
|
WB
Use at an assay dependent concentration. |
Images
-
Western blot - Human B2M (beta 2 Microglobulin) knockout HEK-293T cell line (ab266828)All lanes : Anti-beta 2 Microglobulin antibody [EP2978Y] (ab75853) at 1/5000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : B2M knockout HEK-293T cell lysate
Lane 3 : Wild-type A431 cell lysate
Lane 4 : B2M knockout A431 cell lysate
Performed under reducing conditions.
Observed band size: 12 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-beta 2 Microglobulin antibody [EP2978Y] staining at 1/5000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab75853 was shown to bind specifically to beta 2 Microglobulin. A band was observed at 12 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in B2M knockout cell line ab266828 (knockout cell lysate ab256845). To generate this image, wild-type and B2M 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 fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4°C. 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.
-
Western blot - Human B2M (beta 2 Microglobulin) knockout HEK-293T cell line (ab266828)All lanes : Anti-beta 2 Microglobulin antibody [EPR21752-214] (ab218230) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : B2M knockout HEK-293T cell lysate
Lane 3 : Wild-type A431 cell lysate
Lane 4 : B2M knockout A431 cell lysate
Performed under reducing conditions.
Observed band size: 12 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-beta 2 Microglobulin antibody [EPR21752-214] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab218230 was shown to bind specifically to beta 2 Microglobulin. A band was observed at 12 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in B2M knockout cell line ab266828 (knockout cell lysate ab256845). To generate this image, wild-type and B2M 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 fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4°C. 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.
-
Sanger Sequencing - Human B2M (beta 2 Microglobulin) knockout HEK-293T cell line (ab266828)Sequencing chromatogram displaying sequence edit in exon 1 -
Sanger Sequencing - Human B2M knockout HEK293T cell line (ab266828)Homozygous: 1 bp insertion in exon 1 -
Cell Culture - Human B2M (beta 2 Microglobulin) knockout HEK293T cell line (ab266828)Representative images of B2M knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS XL Core microscope.
Protocols
Datasheets and documents
-
SDS download
-
Datasheet download
References (0)
ab266828 has not yet been referenced specifically in any publications.