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AB257798

Human WHSC1 (NSD2) knockout HeLa cell lysate

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NSD2 KO cell lysate available now. KO validated. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2.
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Sanger Sequencing - Human WHSC1 (NSD2) knockout HeLa cell lysate (AB257798)
  • Sanger seq

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Sanger Sequencing - Human WHSC1 (NSD2) knockout HeLa cell lysate (AB257798)

Homozygous : 1 bp insertion in exon 2

Western blot - Human WHSC1 (NSD2) knockout HeLa cell lysate (AB257798)
  • WB

Lab

Western blot - Human WHSC1 (NSD2) knockout HeLa cell lysate (AB257798)

Lane 1 : Wild-type HeLa cell lysate (20μg)
Lane 2 : WHSC1 knockout HeLa cell lysate (20μg)
Lanes 1- 2 : Merged signal (red and green). Green - ab75359 observed at 80 kDa. Red - loading control, ab181602 observed at 37 kDa.
ab75359 Anti-WHSC1/NSD2 antibody [29D1] - ChIP Grade was shown to specifically react with WHSC1/NSD2 in wild-type HeLa cells in western blot. The band observed in the knockout cell line ab264936 (knockout cell lysate ab257798) lane below 80kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type and WHSC1/NSD2 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. ab75359 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 5 and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-WHSC1/NSD2 antibody [29D1] (<a href='/en-us/products/primary-antibodies/whsc1-nsd2-antibody-29d1-ab75359'>ab75359</a>) at 5 µg/mL

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human WHSC1 (NSD2) knockout HeLa cell lysate (ab257798) at 20 µg

Predicted band size: 152 kDa

Observed band size: 80 kDa

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Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Knockout validation

Sanger Sequencing

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2.

Disease

Adenocarcinoma

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Complementary Products

Primary Antibodies

AB75359

Anti-WHSC1/NSD2 antibody [29D1]

Trial Size

Western blot - Anti-WHSC1/NSD2 antibody [29D1] (AB75359)

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Proteins & Peptides

AB198120

Recombinant human WHSC1/NSD2 protein (Catalytic domain) (GST tag N-Terminus)

Functional Studies - Recombinant human WHSC1/NSD2 protein (Catalytic domain) (GST tag N-Terminus) (AB198120)

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Primary Antibodies

AB239358

Anti-KMT4 / Dot1L antibody [EPR22936-138]

BOND RX™ Validated|RabMAb|Recombinant|Advanced Validation

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KMT4 / Dot1L antibody [EPR22936-138] (AB239358)

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Primary Antibodies

AB300518

Anti-KMT5A / SETD8 / Pr-SET7 antibody [EPR24381-24] - BSA and Azide free

RabMAb|Recombinant|KO Validated

Immunoprecipitation - Anti-KMT5A / SETD8 / Pr-SET7 antibody [EPR24381-24] - BSA and Azide free (AB300518)

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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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Product details

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, 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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What's included?

{ "values": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab257798-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human WHSC1 knockout HeLa cell lysate", "number":"AB257798-CMP01", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human wild-type HeLa cell lysate", "number":"AB257798-CMP02", "productcode":"" } ] } } }
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Properties and storage information

Gene name
NSD2
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing
Zygosity
Homozygous
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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Supplementary information

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

WHSC1 also known as NSD2 is a histone methyltransferase enzyme involved in the methylation of lysine 36 on histone H3. This enzyme facilitates the addition of a methyl group which affects chromatin structure and gene expression. NSD2 is also called WHSC1L1 or Wolf-Hirschhorn syndrome candidate 1-like 1. It has a molecular weight around 153 kDa. Researchers observe the expression of WHSC1/NSD2 predominantly in the nucleus but it is present in various tissues including lymphoid organs and certain types of cancer cells.
Biological function summary

WHSC1/NSD2 functions as an epigenetic regulator modifying histones to modulate transcriptional activity. It exists as part of a larger protein complex that includes several chromatin-associated proteins. Its role in histone methylation influences gene expression impacting cell differentiation survival and proliferation. Through controlling gene transcription WHSC1/NSD2 plays a significant role in development and cellular responses to environmental signals.

Pathways

WHSC1/NSD2 participates in key biological processes such as the cell cycle and DNA damage response pathways. It influences histone modification pathways by interacting with other proteins like SETD2 and H3K36me3 which are important for maintaining genomic integrity. In the DNA damage response NSD2 works together with proteins such as ATM coordinating repair mechanisms and cell cycle progression.

Alterations in WHSC1/NSD2 expression or function associate with multiple myeloma and Wolf-Hirschhorn syndrome. In multiple myeloma overexpression or mutations of WHSC1/NSD2 lead to altered gene expression profiles and promote cancer cell proliferation. Within this context NSD2 often interacts with MMSET and FGFR3 proteins contributing to oncogenic transformation. Additionally mutations in WHSC1/NSD2 link to Wolf-Hirschhorn syndrome a genetic condition characterized by growth delay and intellectual disability.
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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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Product protocols

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Product promise

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For full details, please see our Terms & Conditions

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