AB266828

Human B2M (beta 2 Microglobulin) knockout HEK-293T cell line

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B2M KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1.

We also offer B2M KO in the following cell lines: A549 (ab286573), A-431 (ab261893) and Hep G2 (ab262325)

別名を表示する

B2M, B2MG_HUMAN, Beta 2 microglobin, Beta 2 microglobulin, Beta 2 microglobulin precursor, Beta chain of MHC class I molecules, Beta chain of mhc class 1 proteins, Beta-2-microglobulin form pI 5.3, CDABP0092, Hdcma22p, IMD43

5 Images

Lab

Western blot - Human B2M (beta 2 Microglobulin) knockout HEK-293T cell line (AB266828)

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 A431 and HEK-293T cell lysates with no signal observed at this size in B2M (beta 2 Microglobulin) knockout A-431 cell line ab261893 (knockout cell lysate ab261702) or B2M knockout HEK-293T cell line ab266828 (knockout cell lysate ab256845). To generate this image, wild-type and B2M knockout A431 and HEK-293T cell lysates were analysed. Nitrocellulose membranes were blocked in fluorescent western blot (TBS-based) blocking solution in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed in TBS-T, incubated with secondary antibodies Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20000 dilution for 1 h at room temperature, washed again then imaged.

All lanes:

Western blot - Anti-beta 2 Microglobulin antibody [EP2978Y] (<a href='/products/primary-antibodies/beta-2-microglobulin-antibody-ep2978y-ab75853'>ab75853</a>) at 1/5000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human B2M (beta 2 Microglobulin) knockout HEK-293T cell lysate (<a href='/products/cell-lysates/human-b2m-beta-2-microglobulin-knockout-hek-293t-cell-lysate-ab256845'>ab256845</a>) at 20 µg

Lane 3:

Wild-type A431 cell lysate at 20 µg

Lane 4:

Western blot - Human B2M (beta 2 Microglobulin) knockout A-431 cell lysate (<a href='/products/cell-lysates/human-b2m-beta-2-microglobulin-knockout-a-431-cell-lysate-ab261702'>ab261702</a>) at 20 µg

Predicted band size: 14 kDa

Observed band size: 12 kDa

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Lab

Western blot - Human B2M (beta 2 Microglobulin) knockout HEK-293T cell line (AB266828)

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°. 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.

All lanes:

Western blot - Anti-beta 2 Microglobulin antibody [EPR21752-214] (<a href='/products/primary-antibodies/beta-2-microglobulin-antibody-epr21752-214-ab218230'>ab218230</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

B2M knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human B2M (beta 2 Microglobulin) knockout HEK-293T cell line (ab266828)

Lane 3:

Wild-type A431 cell lysate at 20 µg

Lane 4:

B2M knockout A431 cell lysate at 20 µg

Predicted band size: 14 kDa

Observed band size: 12 kDa

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Sanger Sequencing - Human B2M (beta 2 Microglobulin) knockout HEK-293T cell line (AB266828)

Sanger seq

Unknown

Sanger Sequencing - Human B2M (beta 2 Microglobulin) knockout HEK-293T cell line (AB266828)

Homozygous : 1 bp insertion in exon 1

Cell Culture - Human B2M (beta 2 Microglobulin) knockout HEK-293T cell line (AB266828)

Cell Culture

Unknown

Cell Culture - Human B2M (beta 2 Microglobulin) knockout HEK-293T 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.

Sanger seq

Lab

Sanger Sequencing - Human B2M (beta 2 Microglobulin) knockout HEK-293T cell line (AB266828)

Sequencing chromatogram displaying sequence edit in exon 1.

Key facts

細胞タイプ

HEK-293T

生物種

Human

組織

Kidney

製品の状態

Liquid

form

ノックアウト検証方法

Sanger Sequencing

ノックアウト変異

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"" }, "Sanger seq": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"" }, "Cell Culture": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"" } } }

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製品の詳細

Red fluorescence is observed in this KO cell line when using imaging based assays, the cause of the fluorescence is not tested.

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.

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製品内容

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出荷温度及び保存条件

遺伝子名

B2M

遺伝子編集のタイプ

Knockout

遺伝子編集の方法

CRISPR technology

ノックアウト検証方法

Sanger Sequencing

接合型

Homozygous

出荷温度

Dry Ice

短期保存温度

-196°C

長期保存温度

-196°C

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取り扱い方法

初回取り扱いガイドライン

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.

継代培養ガイドライン

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.

培養培地

DMEM (High Glucose) + 10% FBS

凍結保存培地

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

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補足情報

This supplementary information is collated from multiple sources and compiled automatically.

Beta-2-Microglobulin (B2M) is a component of the class I major histocompatibility complex (MHC I) and plays an important role in presenting peptides to the immune system. B2M weighs approximately 11.8 kDa and is found abundantly in all nucleated cells. It has alternate names such as B2 microglobulin or beta-2-microglobulin. This protein is present in the cell membrane as a part of the MHC I which is important for immune surveillance. Additionally B2M is detectable in various biological fluids including serum and its levels can reflect physiological and pathological states.

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.

Beta-2-microglobulin is associated with conditions such as beta-2-microglobulin amyloidosis and certain lymphoproliferative disorders. Elevated levels of B2M in serum serve as a marker for diseases like multiple myeloma where the protein level correlates with disease severity. B2M-related amyloidosis frequently occurs in patients undergoing long-term dialysis where amyloid deposits accumulate in tissues. Linking B2M to immune system dysfunction studies have shown interactions with other proteins including components of the immune system like HLA-A and HLA-B highlighting B2M's relevance in diagnosing and understanding these conditions.

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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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製品プロトコール

Visit the General protocols
Visit the Troubleshooting

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ターゲットの情報

See full target information B2M

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Human B2M (beta 2 Microglobulin) knockout HEK-293T cell line