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AB257247

Human NFKB2 (NFkB p100/NFKB2) knockout Hep G2 cell lysate

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NFKB2 KO cell lysate available now. KO validated by. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 28 bp deletion in exon12.

View Alternative Names

CVID10, DNA-binding factor KBF2, H2TF1, LYT 10, Lymphocyte translocation chromosome 10 protein, NFKB p52/p100 subunit, Nuclear factor Kappa B subunit 2, Nuclear factor of kappa light polypeptide gene enhancer in B cells 2 (p49/p100), Nuclear factor of kappa light polypeptide gene enhancer in B-cells 2, Oncogene Lyt-10, Transcription factor NFKB2

4 Images
Western blot - Human NFKB2 (NFkB p100/NFKB2) knockout Hep G2 cell lysate (AB257247)
  • WB

Lab

Western blot - Human NFKB2 (NFkB p100/NFKB2) knockout Hep G2 cell lysate (AB257247)

Lane 1 : Wild-type HepG2 cell lysate (20μg)
Lane 2 : NFKB2 knockout HepG2 cell lysate (20μg)
Lanes 1- 2 : Merged signal (red and green). Green - ab109440 observed at 120 kDa. Red - loading control, ab8245 observed at 37 kDa.
ab109440 Anti-NFkB p100/NFKB2 antibody [EPR4686] was shown to specifically react with NFkB p100/NFKB2 in wild-type HepG2 cells in western blot. The band observed in the knockout cell line ab262323 (knockout cell lysate ab257247) lane below 97kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type and NFkB p100/NFKB2 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. ab109440 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) 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-NFkB p100/NFKB2 antibody [EPR4686] (<a href='/en-us/products/primary-antibodies/nfkb-p100-nfkb2-antibody-epr4686-ab109440'>ab109440</a>) at 1/1000 dilution

Lane 1:

Wild-type HepG2 cell lysate at 20 µg

Lane 2:

NFKB2 knockout HeLa cell lysate at 20 µg

Predicted band size: 97 kDa

Observed band size: 120 kDa

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Sanger Sequencing - Human NFKB2 (NFkB p100/NFKB2) knockout Hep G2 cell lysate (AB257247)
  • Sanger seq

Unknown

Sanger Sequencing - Human NFKB2 (NFkB p100/NFKB2) knockout Hep G2 cell lysate (AB257247)

Homozygous : 28 bp deletion in exon12

Western blot - Human NFKB2 (NFkB p100/NFKB2) knockout Hep G2 cell lysate (AB257247)
  • WB

Lab

Western blot - Human NFKB2 (NFkB p100/NFKB2) knockout Hep G2 cell lysate (AB257247)

Lane 1 : Wild-type HepG2 cell lysate (20μg)
Lane 2 : NFKB2 knockout HepG2 cell lysate (20μg)
Lanes 1- 2 : Merged signal (red and green). Green - ab175192 observed at 120 kDa. Red - loading control, ab8245 observed at 37 kDa.
ab175192 Anti-NFkB p100/NFKB2 antibody [EPR4686-66] was shown to specifically react with NFkB p100/NFKB2 in wild-type HepG2 cells in western blot. The band observed in the knockout cell line ab262323 (knockout cell lysate ab257247) lane below 97kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type and NFkB p100/NFKB2 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. ab175192 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 10000 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-NFkB p100/NFKB2 antibody [EPR4686-66] (<a href='/en-us/products/primary-antibodies/nfkb-p100-nfkb2-antibody-epr4686-66-ab175192'>ab175192</a>) at 1/1000 dilution

Lane 1:

Wild-type HepG2 cell lysate at 20 µg

Lane 2:

Western blot - Human NFKB2 (NFkB p100/NFKB2) knockout Hep G2 cell lysate (ab257247)

Predicted band size: 97 kDa

Observed band size: 120 kDa

false

Western blot - Human NFKB2 (NFkB p100/NFKB2) knockout Hep G2 cell lysate (AB257247)
  • WB

Lab

Western blot - Human NFKB2 (NFkB p100/NFKB2) knockout Hep G2 cell lysate (AB257247)

All lanes:

Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686] (<a href='/en-us/products/primary-antibodies/nfkb-p100-nfkb2-antibody-epr4686-ab109440'>ab109440</a>) at 1/1000 dilution

Lane 1:

Wild-type HepG2 cell lysate at 20 µg

Lane 2:

Western blot - Human NFKB2 (NFkB p100/NFKB2) knockout Hep G2 cell lysate (ab257247) at 20 µg

Secondary

Lanes 1 - 2:

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 1/20000 dilution

Lanes 1 - 2:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution

Predicted band size: 30 kDa,97 kDa

Observed band size: 120 kDa,42 kDa,37 kDa

false

Key facts

Cell type

Hep G2

Species or organism

Human

Tissue

Liver

Knockout validation

Sanger Sequencing

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 28 bp deletion in exon12.

Disease

Hepatocellular Carcinoma

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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 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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What's included?

{ "values": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab257247-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human wild-type HepG2 cell lysate", "number":"AB257247-CMP01", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human NFKB2 knockout HepG2 cell lysate", "number":"AB257247-CMP02", "productcode":"" } ] } } }
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Properties and storage information

Gene name
NFKB2
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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Quality control

STR analysis

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

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Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Male

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Product protocols

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Complementary Products

Primary Antibodies

AB175192

Anti-NFkB p100/NFKB2 antibody [EPR4686-66]

RabMAb|Recombinant|KO Validated

Immunoprecipitation - Anti-NFkB p100/NFKB2 antibody [EPR4686-66] (AB175192)

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Primary Antibodies

AB76302

Anti-NF-kB p65 (phospho S536) antibody [EP2294Y]

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Immunoprecipitation - Anti-NF-kB p65 (phospho S536) antibody [EP2294Y] (AB76302)

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Multiplex Kits

AB228529

NF-kB Signaling Pathway Antibody Sampler Panel

Recombinant

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - NF-kB Signaling Pathway Antibody Sampler Panel (AB228529)

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Assay Kits

AB133112

NFkB p65 Transcription Factor Assay Kit

Functional Studies - NFkB p65 Transcription Factor Assay Kit (AB133112)

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Product promise

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For full details, please see our Terms & Conditions

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