AB257463

Human H2AFY (mH2A1) knockout HEK-293T cell lysate

Name: Human H2AFY (mH2A1) knockout HEK-293T cell lysate (ab257463) | Abcam
Brand: Abcam Limited
SKU: AB257463
Price: 111000 JPY
Availability: InStock

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MACROH2A1 KO cell lysate available now. KO validated. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 13 bp deletion in exon 2.

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Lab

Western blot - Human H2AFY (mH2A1) knockout HEK-293T cell lysate (AB257463)

Lane 1 : Wild-type HEK-293T cell lysate (20µg)

Lane 2 : H2AFY knockout HEK-293T cell lysate (20µg)

Lanes 1- 2 : Merged signal (red and green). Green - ab183041 observed at 40 kDa. Red - loading control, ab7291 observed at 50 kDa.

ab183041 Anti-mH2A1 antibody [EPR9359(2)] was shown to specifically react with mH2A1 in wild-type HEK-293T cells in western blot. The band observed in the knockout cell line ab266241 (knockout cell lysate ab257463) lane below 40kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type and mH2A1 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab183041 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4^°C at 1 in 1000 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-mH2A1 antibody [EPR9359(2)] (<a href='/products/primary-antibodies/mh2a1-antibody-epr93592-ab183041'>ab183041</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

H2AFY knockout HEK-293T cell lysate at 20 µg

Predicted band size: 40 kDa

Observed band size: 40 kDa

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Sanger Sequencing - Human H2AFY (mH2A1) knockout HEK-293T cell lysate (AB257463)

Sanger seq

Unknown

Sanger Sequencing - Human H2AFY (mH2A1) knockout HEK-293T cell lysate (AB257463)

Homozygous : 13 bp deletion in exon 2

Key facts

細胞タイプ

HEK-293T

生物種

Human

組織

Kidney

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: 13 bp deletion in exon 2.

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p>Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.</p>" } } }

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製品の詳細

Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

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.

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製品内容

{ "values": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab257463-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human H2AFY knockout HEK293T cell lysate", "number":"AB257463-CMP01", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human wild-type HEK293T cell lysate", "number":"AB257463-CMP02", "productcode":"" } ] } } }

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出荷温度及び保存条件

遺伝子名

MACROH2A1

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

Ambient - Can Ship with Ice

短期保存温度

-20°C

長期保存温度

-20°C

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補足情報

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

MH2A1 also known as macroH2A1 is a histone variant with a distinguished structure characterized by an extended C-terminal macro domain. It has a molecular mass of approximately 42-45 kDa. This histone variant is widely expressed across various tissues showing particularly high levels in differentiated cells. mH2A1 incorporates into chromatin impacting nucleosome stability and chromatin dynamics. Researchers have also identified an isoform known as H2AFY which further highlights the structural complexity and versatile roles of mH2A1 in cellular regulation.

Biological function summary

MH2A1 plays significant roles in chromatin remodeling and gene expression regulation. It is part of the nucleosome replacing canonical histone H2A and influencing the accessibility of chromatin to transcriptional machinery. Through this function mH2A1 can repress or activate gene expression depending on cellular context and interacting partners. Its incorporation into chromatin is associated with the silencing of certain genomic regions such as inactive X chromosome in females with potential implications for epigenetic modifications and cellular differentiation.

Pathways

MH2A1 is notably involved in pathways regulating chromatin organization and cell differentiation. It plays a role in the Polycomb Repressive Complex 2 (PRC2) pathway affecting histone methylation and gene silencing. Furthermore mH2A1 interacts with proteins like EZH2 in PRC2 which carry out methylation at histone H3 on lysine 27. These interactions highlight mH2A1's involvement in maintaining repressive chromatin states and controlling gene expression patterns important for normal cell function.

MH2A1 is connected with cancer and metabolic diseases. Its altered expression has been observed in various cancers where it might impact tumor progression by modulating genes linked to proliferation and differentiation. mH2A1 influences pathways shared with proteins like BRAC1 that are involved in DNA repair processes. Additionally changes in mH2A1 expression are linked to metabolic disorders especially those affecting lipid metabolism and insulin responses suggesting its potential role in regulating metabolic pathways and influencing disease progression.

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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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Abcam product promise

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Human H2AFY (mH2A1) knockout HEK-293T cell lysate