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AB257482

Human INA (alpha Internexin) knockout HeLa cell lysate

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INA KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 179 bp deletion in exon1 and 1 bp insertion in exon1.
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Western blot - Human INA (alpha Internexin) knockout HeLa cell lysate (AB257482)
  • WB

Lab

Western blot - Human INA (alpha Internexin) knockout HeLa cell lysate (AB257482)

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

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

Lane 3 : SH-SY5Y cell lysate (20 μg)

Lanes 1-4 : Merged signal (red and green). Green - ab40758 observed at 66 kDa. Red - loading control ab8245 observed at 36 kDa.

ab40758 Anti-alpha Internexin antibody [EP676Y] was shown to specifically react with alpha Internexin in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265264 (knockout cell lysate ab257482) was used. Wild-type and alpha Internexin knockout samples were subjected to SDS-PAGE. ab40758 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) 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-alpha Internexin antibody [EP676Y] (<a href='/en-us/products/primary-antibodies/alpha-internexin-antibody-ep676y-ab40758'>ab40758</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

INA knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human INA (alpha Internexin) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-ina-alpha-internexin-knockout-hela-cell-line-ab265264'>ab265264</a>)

Lane 3:

SH-SY5Y 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 1/10000 dilution

Predicted band size: 52 kDa,55 kDa

Observed band size: 60 kDa,66 kDa

false

Sanger Sequencing - Human INA (alpha Internexin) knockout HeLa cell lysate (AB257482)
  • Sanger seq

Unknown

Sanger Sequencing - Human INA (alpha Internexin) knockout HeLa cell lysate (AB257482)

Allele-1 : 1 bp insertion in exon1

Sanger Sequencing - Human INA (alpha Internexin) knockout HeLa cell lysate (AB257482)
  • Sanger seq

Unknown

Sanger Sequencing - Human INA (alpha Internexin) knockout HeLa cell lysate (AB257482)

Allele-2 : 179 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, 179 bp deletion in exon1 and 1 bp insertion in exon1.

Disease

Adenocarcinoma

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Complementary Products

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Recombinant Human alpha Internexin protein

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Primary Antibodies

AB40758

Anti-alpha Internexin antibody [EP676Y]

RabMAb|Recombinant|KO Validated

Western blot - Anti-alpha Internexin antibody [EP676Y] (AB40758)

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Cell Lines & Lysates

AB265264

Human INA (alpha Internexin) knockout HeLa cell line

Advanced Validation

Western blot - Human INA (alpha Internexin) knockout HeLa cell line (AB265264)

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Reactivity data

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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, 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?

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Properties and storage information

Gene name
INA
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.

Alpha Internexin also known as alpha-Int or INA is a neuronal intermediate filament protein with a molecular mass of approximately 66 kDa. It plays a significant role in the formation of the neuronal cytoskeleton providing structural support to axons. Alpha Internexin is expressed mainly in the central nervous system and is found in neurons throughout the brain and spinal cord. Despite sharing functions with other intermediate filament proteins like NF-L alpha Internexin is unique in its distribution and timing of expression during neural development.
Biological function summary

Alpha Internexin contributes to the composition of the neurofilament complex which includes proteins like NF-M NF-L and NF-H. This complex stabilizes neuronal architecture and ensures the integrity of the axonal structure. Presence of alpha Internexin is important for maintaining the dynamic processes of axonal transport and neuron stability. It acts as a scaffolding that supports cellular transport mechanisms important for neuron function and survival especially in developing neurons where it leads the polymerization of other neurofilaments.

Pathways

Alpha Internexin interacts within the neurofilament network a major scaffolding component of neurons. It associates with microtubule-associated proteins influencing axonal growth and stabilization pathways. Moreover it plays a part in the MAPK (mitogen-activated protein kinases) signaling pathways which regulate diverse cellular activities such as proliferation and differentiation. Its interactions with related proteins such as plectin and dynein highlight its role in crosslinking cytoskeletal components and facilitating intracellular transport.

Alpha Internexin has been implicated in neurodegenerative conditions such as Amyotrophic Lateral Sclerosis (ALS) and Alzheimer's disease. Misregulation or mutation in alpha Internexin can contribute to the pathological hallmarks of these diseases including protein aggregation and neuronal death. In ALS the presence of alpha Internexin inclusions correlates with disease pathology alongside other neurofilament proteins like NF-L. In Alzheimer's disease abnormal phosphorylation and aggregation of alpha Internexin are associated with neurofibrillary tangles which are indicative of neuronal injury and loss.
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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

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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