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AB258533

Human NF1 (Neurofibromin) knockout HeLa cell lysate

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NF1 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon1.
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Western blot - Human NF1 (Neurofibromin) knockout HeLa cell lysate (AB258533)
  • WB

Lab

Western blot - Human NF1 (Neurofibromin) knockout HeLa cell lysate (AB258533)

Lane 1 : Wild-type HeLa cell lysate (20μg)

Lane 2 : NF1 knockout HeLa cell lysate (20μg)

Lane 3 : HAP1 cell lysate (20μg)

Lanes 1-3 : Merged signal (red and green). Green - ab238142 observed at 319 kDa. Red - loading control ab7291 observed at 50 kDa.

ab238142 Recombinant Anti-NF1 antibody [EPR22989-68] was shown to specifically react with NFIB in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264725 (knockout cell lysate ab258533) was used. Wild-type and NF1 knockout samples were subjected to SDS-PAGE. ab238142 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 1000 dilution 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-Neurofibromin antibody [EPR22989-68] (<a href='/en-us/products/primary-antibodies/neurofibromin-antibody-epr22989-68-ab238142'>ab238142</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

NFIB knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human NF1 (Neurofibromin) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-nf1-neurofibromin-knockout-hela-cell-line-ab264725'>ab264725</a>)

Lane 3:

HAP1 cell lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 319 kDa

Observed band size: 319 kDa

false

Sanger Sequencing - Human NF1 (Neurofibromin) knockout HeLa cell lysate (AB258533)
  • Sanger seq

Unknown

Sanger Sequencing - Human NF1 (Neurofibromin) knockout HeLa cell lysate (AB258533)

Homozygous : 1 bp deletion in exon1

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon1.

Disease

Adenocarcinoma

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Primary Antibodies

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RabMAb|Recombinant|KO Validated

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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></p>" } } }
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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

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?

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Properties and storage information

Gene name
NF1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
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.

Neurofibromin is a protein that functions as a Ras GTPase-activating protein acting to promote the hydrolysis of Ras-bound GTP to GDP which inactivates Ras signaling. This regulatory function involves the suppression of excessive cell proliferation and differentiation. Alternate names for neurofibromin include NF1 or neurofibromin 1. The mass of neurofibromin is approximately 327 kDa. Neurofibromin is expressed widely across various tissues with notable expression in the nervous system and the brain.
Biological function summary

Neurofibromin regulates pathways involved in cell growth and development. It does not form part of a large multiprotein complex but interacts with microtubules and other signaling proteins to carry out its regulatory functions. Neurofibromin primarily affects pathways that manage cell cycle progression and it plays a critical role in inhibiting uncontrolled cell growth ensuring proper cellular homeostasis.

Pathways

Neurofibromin fits into the Ras-MAPK pathway where it controls the activity of Ras an important regulatory protein involved in signal transduction related to cell division and differentiation. Neurofibromin's action on Ras directly impacts the activation of the MAPK pathway which is important in mediating cellular responses to growth signals. This protein also interacts functionally with proteins like SOS1 another regulator of Ras emphasizing its role in fine-tuning signal cascades that ensure balanced cellular function.

Neurofibromin's dysregulation is most commonly associated with neurofibromatosis type 1 a genetic disorder characterized by the development of benign tumors in the nervous system. The loss of neurofibromin function leads to unregulated Ras activity resulting in tumorigenesis. Additionally neurofibromin is implicated in certain sporadic cancers due to its role in controlling cell proliferation. In neurofibromatosis type 1 interactions between neurofibromin and proteins such as p53 a known tumor suppressor highlight its significance in disease pathways and tumor suppression mechanisms.
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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

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

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