Recombinant Anti-alpha Internexin antibody [EP676Y] (ab40758)
![Western blot - Anti-alpha Internexin antibody [EP676Y] (ab40758)](https://www.abcam.com/ps/products/40/ab40758/Images/ab40758-391341-anti-alpha-internexin-antibody-ep676y-western-blot-human-knockout-lysate.jpg "Western blot - Anti-alpha Internexin antibody [EP676Y] (ab40758)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP676Y] to alpha Internexin
- Suitable for: WB, IP, IHC-FoFr, Flow Cyt, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-alpha Internexin antibody [EP676Y]
See all alpha Internexin primary antibodies -
Description
Rabbit monoclonal [EP676Y] to alpha Internexin -
Host species
Rabbit -
Tested applications
Suitable for: WB, IP, IHC-FoFr, Flow Cyt, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Human
Does not react with: Apteronotus leptorhynchus -
Immunogen
Synthetic peptide within Human alpha Internexin aa 400-500. The exact sequence is proprietary.
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Positive control
- HeLa, SHSY5Y cell lysate, human brain lysate.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP676Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab40758 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB | (2) |
1/5000 - 1/10000. Detects a band of approximately 60 kDa (predicted molecular weight: 66 kDa).
|
| IP |
Use at an assay dependent concentration.
|
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| IHC-FoFr | (1) |
Use at an assay dependent concentration.
|
| Flow Cyt |
Use at an assay dependent concentration.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
|
| IHC-P | (1) |
1/250 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
|
| ICC/IF | (1) |
1/250 - 1/500.
|
| Notes |
|---|
|
WB
1/5000 - 1/10000. Detects a band of approximately 60 kDa (predicted molecular weight: 66 kDa). |
|
IP
Use at an assay dependent concentration. |
|
IHC-FoFr
Use at an assay dependent concentration. |
|
Flow Cyt
Use at an assay dependent concentration. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
|
IHC-P
1/250 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
|
ICC/IF
1/250 - 1/500. |
Target
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Function
Class-IV neuronal intermediate filament that is able to self-assemble. It is involved in the morphogenesis of neurons. It may form an independent structural network without the involvement of other neurofilaments or it may cooperate with NF-L to form the filamentous backbone to which NF-M and NF-H attach to form the cross-bridges. -
Tissue specificity
Found predominantly in adult CNS. -
Sequence similarities
Belongs to the intermediate filament family. -
Developmental stage
Expressed in brain as early as the 16th week of gestation, and increased rapidly and reached a steady state level by the 18th week of gestation. -
Post-translational
modificationsO-glycosylated.
Phosphorylated upon DNA damage, probably by ATM or ATR. - Information by UniProt
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Database links
- Entrez Gene: 9118 Human
- Entrez Gene: 226180 Mouse
- Entrez Gene: 24503 Rat
- Omim: 605338 Human
- SwissProt: Q16352 Human
- SwissProt: P46660 Mouse
- SwissProt: P23565 Rat
- Unigene: 500916 Human
see all -
Alternative names
- 66 kDa neurofilament protein antibody
- AINX_HUMAN antibody
- Alpha Inx antibody
see all
Images
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Western blot - Anti-alpha Internexin antibody [EP676Y] (ab40758)All lanes : Anti-alpha Internexin antibody [EP676Y] (ab40758) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : INA knockout HeLa cell lysate
Lane 3 : SH-SY5Y cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 66 kDa
Observed band size: 66 kDaLanes 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.
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![Western blot - Anti-alpha Internexin antibody [EP676Y] (ab40758)](https://www.abcam.com/ps/products/40/ab40758/Images/40758a.jpg)
Western blot - Anti-alpha Internexin antibody [EP676Y] (ab40758)Anti-alpha Internexin antibody [EP676Y] (ab40758) at 1/10000 dilution + SHSY5Y cell lysate at 10 µg
Predicted band size: 66 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted? -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha Internexin antibody [EP676Y] (ab40758)](https://www.abcam.com/ps/products/40/ab40758/Images/ab40758-53864-anti-alpha-internexin-antibody-ep676y-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha Internexin antibody [EP676Y] (ab40758)Positive staining of alpha Internexin on paraffin-embedded human brain tissue (A) using ab40758 diluited 1:250. Negative staining of paraffin-embedded human colon (B), Kidney (C) and uterus (D) tissues using ab40758 under the same conditions.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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![Immunocytochemistry/ Immunofluorescence - Anti-alpha Internexin antibody [EP676Y] (ab40758)](https://www.abcam.com/ps/products/40/ab40758/Images/ab40758-227989-ab40758iccif.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-alpha Internexin antibody [EP676Y] (ab40758)This image is courtesy of an anonymous Abreviewab40758 staining alpha Internexin in rat glial cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with saponin and blocked with 1% BSA for 10 minutes at 25°C. Samples were incubated with primary antibody (1/200) for 10 hours at 4°C. An undiluted Alexa Fluor® 594-conjugated donkey anti-rabbit IgG polyclonal was used as the secondary antibody.
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![Immunocytochemistry/ Immunofluorescence - Anti-alpha Internexin antibody [EP676Y] (ab40758)](https://www.abcam.com/ps/products/40/ab40758/Images/40758c.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-alpha Internexin antibody [EP676Y] (ab40758)Immunofluorescent staining of alpha Internexin on PC 12 cells using ab40758 diluted 1:250. -
![Anti-alpha Internexin antibody [EP676Y] (ab40758)](https://www.abcam.com/ps/products/40/ab40758/Images/ab40758-7-benefits-of-recombinant-antibodies.png)
Anti-alpha Internexin antibody [EP676Y] (ab40758)
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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Datasheet download
References (7)
ab40758 has been referenced in 7 publications.
- Morandell J et al. Cul3 regulates cytoskeleton protein homeostasis and cell migration during a critical window of brain development. Nat Commun 12:3058 (2021). PubMed: 34031387
- Iqbal J et al. Selenium positively affects the proteome of 3 × Tg-AD mice cortex by altering the expression of various key proteins: unveiling the mechanistic role of selenium in AD prevention. J Neurosci Res 96:1798-1815 (2018). PubMed: 30054946
- Armao D et al. Autonomic nervous system involvement in the giant axonal neuropathy (GAN) KO mouse: implications for human disease. Clin Auton Res 26:307-13 (2016). PubMed: 27369358
- Ayyadevara S et al. Proteins that mediate protein aggregation and cytotoxicity distinguish Alzheimer's hippocampus from normal controls. Aging Cell 15:924-39 (2016). IHC-P ; Human . PubMed: 27448508
- Xing R et al. Spatial memory impairment by TRPC1 depletion is ameliorated by environmental enrichment. Oncotarget 7:27855-73 (2016). WB . PubMed: 27034165
- Coffey RT et al. Ubiquilin-mediated Small Molecule Inhibition of Mammalian Target of Rapamycin Complex 1 (mTORC1) Signaling. J Biol Chem 291:5221-33 (2016). PubMed: 26740621
- Louis ED et al. Neurofilament protein levels: quantitative analysis in essential tremor cerebellar cortex. Neurosci Lett 518:49-54 (2012). PubMed: 22561033
