Human INA (alpha Internexin) knockout HeLa cell line
- Advanced Validation
- What is this?
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INA KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 179 bp deletion in exon 1 and 1 bp insertion in exon 1. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
View Alternative Names
66 kDa neurofilament protein, AINX_HUMAN, Alpha-Inx, Alpha-internexin, INA, Internexin neuronal intermediate filament protein alpha, MGC12702, NEF 5, NF-66, Neurofilament 5, Neurofilament 5 (66kD), Neurofilament 66 tax binding protein, Neurofilament-66, TXBP 1
- WB
Lab
Western blot - Human INA (alpha Internexin) knockout HeLa cell line (AB265264)
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 (ab265264)
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 seq
Unknown
Sanger Sequencing - Human INA (alpha Internexin) knockout HeLa cell line (AB265264)
Allele-1 : 1 bp insertion in exon 1.
- Sanger seq
Unknown
Sanger Sequencing - Human INA (alpha Internexin) knockout HeLa cell line (AB265264)
Allele-2 : 179 bp deletion in exon 1.
Reactivity data
Product details
We will provide viable cells that proliferate on revival.
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Handling procedures
Initial handling guidelines
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
DMEM (High Glucose) + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
Primary Antibodies
AB40758
Anti-alpha Internexin antibody [EP676Y]
RabMAb|Recombinant|KO Validated
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha Internexin antibody [EP676Y] (AB40758)](https://content.abcam.com/products/images/alpha-internexin-antibody-ep676y-ab40758--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img131288.jpg)
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com