SpecificityThe specificity of this antibody has been confirmed by WB. This antibody is specific for the 64-66 kDa alpha-internexin protein. Molecular weight will depend on species. Hu, Rat, Ms, Fel, and other mammals
Species ReactivityCat, Human, Mouse, Other Mammals, Rat
Immunogen DescriptionRecombinant rat alpha-internexin expressed and purified from E. coli
Application DetailsWestern Blotting (WB), Immunocytochemistry (ICC) and Immunohistochemistry on paraffin sections (IHC). The recommended dilution for WB is 1:5000-10,000. The epitope recognised by this antibody is in the C-terminal non-helical extension of the protein and is unusually resistant to aldehyde fixation so this antibody is ideally suited for studies of paraffin embedded formalin fixed histological sections. The recommended dilution for IHC is 1:1,000-5,000. For IC, dilution is 1:250-500. Biosensis recommends optimal dilutions/concentrations should be determined by the end user.
TargetInternexin alpha (Alpha-Inx)
SpecificityThe specificity of this antibody has been confirmed by WB. This antibody is specific for the 64-66 kDa alpha-internexin protein. Molecular weight will depend on species. Hu, Rat, Ms, Fel, and other mammals
Target Host SpeciesRat
Species ReactivityCat, Human, Mouse, Other Mammals, Rat
Antibody HostMouse
Antibody TypeMonoclonal
Antibody IsotypeIgG1
Clone Name1D2
ConjugateUnconjugated
Immunogen DescriptionRecombinant rat alpha-internexin expressed and purified from E. coli
Purity DescriptionProtein G purified
FormatLyophilized from PBS buffer pH 7.2-7.6 with 0.1% trehalose, and sodium azide
Reconstitution InstructionsSpin vial briefly before opening. Reconstitute with 250 µL sterile-filtered, ultrapure water to achieve a 1 mg/mL concentration. Centrifuge to remove any insoluble material.
Storage InstructionsAfter reconstitution of lyophilized antibody, aliquot and store at -20°C for a higher stability. Avoid freeze-thaw cycles.
Batch NumberPlease see item label.
Expiration Date12 months after date of receipt (unopened vial).
Scientific BackgroundNeurofilaments can be defined as the intermediate or 10nm diameter filaments found in neuronal cells. They are composed a mixture of subunits which often includes the neurofilament triplet proteins, NF-L, NF-M and NF-H. Neurofilaments may also include peripherin, alpha-internexin, nestin and in some cases vimentin. Alpha-internexin is a ~66 kDa Class IV intermediate filament subunit expressed in large amounts early in neuronal development, but is downregulated in many neurons as development procedes. Many classes of mature neurons contain alpha-internexin in addition to NF-L, NF-M and NF-H. In some mature neurons alpha-internexin is the only neurofilament subunit expressed. Antibodies to alpha-internexin are therefore unique probes to study and classify neuronal types and follow their processes in sections and in tissue culture. In addition the very early developmental expression of alpha-internexin means its presence is an early and convenient diagnostic feature of neuronal progenitors cells and other cell committed to the neuronal lineage.
Mixed cultures of rat CNS cells stained with Mouse monoclonal antibody to Internexin alpha [1D2] M-1378-250 (red) and Chicken polyclonal antibody to Microtubule associated protein 2 C-1382-50 (green). The internexin alpha is localized primarily in neuronal axons in these cultures, while the perikarya and dendrites of neurons stain strongly for MAP2.
Left: Analysis of alpha-internexin expression in rat cerebellum with mouse antibody to alpha-internexin (1:5,000, green) by Immunohistochemistry. Co-staining was performed with chicken antibody to calretinin (C-1801-50, 1:2,000, red). IHC Method: Following transcardial perfusion of rat with 4% PFA, brain was post-fixed for 24 hours, cut to 45 um, and free-floating sections were stained. The alpha-internexin antibody selectively stains neuronal processes, in particular parallel fibers, the axons of granule cells. Calretinin antibody stains interneurons predominantly in the molecular layer of the cerebellum. Right: Western blot analysis of alpha-internexin expression in tissue lysates using mouse antibody to alpha-internexin (1:10,000, green): [1] protein standard, [2] rat brain, [3] rat spinal cord, [4] mouse brain, [5] mouse spinal cord, [6] pig spinal cord, [7] cow spinal cord. Alpha-internexin protein migrates on the gel with apparent molecular weight of 64 to 66 kDa with slight variability among species.
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