Product NameNeurofilament heavy polypeptide, phosphorylated, (pNF-H), Mouse Monoclonal Antibody (NAP4)
Product DescriptiongoogleMouse anti-Neurofilament heavy polypeptide, phosphorylated, (pNF-H), Monoclonal Antibody (Unconjugated), suitable for WB, Immunostaining and FC.
Alternative NamesNF-H; NFH; NF-200; NF200; NF-H; NEFH; N52; Neurofilament heavy polypeptide; Neurofilament triplet H protein; 200 kDa neurofilament protein; KIAA0845
Application(s)FC, IF, ICC, IHC, WB
Antibody HostMouse
Antibody TypeMonoclonal
SpecificitySpecies cross-reactivity includes human, rat, mouse, cow, pig, horse and chicken. This antibody recognises phosphorylated NF-H KSP (lysine-serine-proline) type sequences. In some species there is some cross-reactivity with the related KSP sequences found in subunit NF-M. Predicted to react with other mammals due to sequence homology.
Species ReactivityBovine, Chicken, Horse, Human, Mouse, Other Mammals (Predicted), Pig, Rat
Immunogen DescriptionThis antibody has been made against a native axonal phosphorylated NF-H purified from bovine spinal cord.
Product DescriptionMouse anti-Neurofilament heavy polypeptide, phosphorylated, (pNF-H), Monoclonal Antibody (Unconjugated), suitable for WB, Immunostaining and FC.
Application DetailsWestern blot (WB), Immunocytochemistry (ICC) / Immunofluorescence (IF), and Immunohistochemistry (IHC) and Flow Cytometry (FC). A dilution of 1:10,000 is recommended for WB. A dilution of 1:1,000 is recommended for ICC/IF and IHC. This antibody recognises NF-H in frozen sections, tissue culture and in formalin-fixed sections. A dilution of 2 ug/10^6 cells is recommended for FC. Biosensis recommends optimal dilutions/concentrations should be determined by the end user.
TargetNeurofilament heavy polypeptide, phosphorylated (pNF-H)
SpecificitySpecies cross-reactivity includes human, rat, mouse, cow, pig, horse and chicken. This antibody recognises phosphorylated NF-H KSP (lysine-serine-proline) type sequences. In some species there is some cross-reactivity with the related KSP sequences found in subunit NF-M. Predicted to react with other mammals due to sequence homology.
Target Host SpeciesBovine
Species ReactivityBovine, Chicken, Horse, Human, Mouse, Other Mammals (Predicted), Pig, Rat
Antibody HostMouse
Antibody TypeMonoclonal
Antibody IsotypeIgG1
Clone NameNAP4
ConjugateUnconjugated
Immunogen DescriptionThis antibody has been made against a native axonal phosphorylated NF-H purified from bovine spinal cord.
Purity DescriptionPurified mouse IgG
FormatLyophilized from PBS buffer pH 7.2-7.6 with 0.1% trehalose, and 5mM sodium azide.
Reconstitution InstructionsSpin vial briefly before opening. Reconstitute with 100 µL sterile-filtered, ultrapure water to achieve a 1 mg/mL concentration. Centrifuge to remove any insoluble material.
Storage InstructionsStore lyophilized antibody at 2-8°C After reconstitution of lyophilized antibody, aliquot and store at -20°C for a higher stability. Avoid freeze-thaw cycles. Store at 4°C for up to one month for short term storage and frequent use.
Batch NumberPlease see item label.
Expiration Date12 months after date of receipt (unopened vial).
Alternative NamesNF-H; NFH; NF-200; NF200; NF-H; NEFH; N52; Neurofilament heavy polypeptide; Neurofilament triplet H protein; 200 kDa neurofilament protein; KIAA0845
Scientific BackgroundNeurofilaments are the 10nm or intermediate filament proteins found specifically in neurons, and are composed predominantly of three major proteins called NF-L, NF-M and NF-H, though other filament proteins may be included also. The major function of neurofilaments is likely to control the diameter of large axons. NF-L is the neurofilament light or low molecular weight polypeptide and runs on SDS-PAGE gels at 68-70kDa with some variability across species. Antibodies to NF-L are useful for identifying neuronal cells and their processes in cell culture and sectioned material. NF-L antibody can also be useful for the visualization of neurofilament rich accumulations seen in many neurological diseases, such as Lou Gehrig’s disease (ALS), giant axon neuropathy, Charcot-Marie Tooth disease and others. (Ref: uniprot.org)
Image of a human cerebellar cortex section by Immunohistochemistry. The section was stained with M-1387-50, Neurofilament heavy polypeptide, phosphorylated (pNF-H), Clone NAP4, Mouse mAb (brown) and co-stained with heamatoxylin-eosin (blue). This antibody stains prominent basket cell axons surrounding the large Purkinje neurons. The granule cell layer is at the bottom of the image with the molecular layer at the top. IHC method: Section are fixed in formalin, embedded in paraffin using the ABC (avidin biotin conjugate).
Analysis by flow cytometry of Neurofilament heavy polypeptide, phosphorlylated (pNF-H), (endogenous) expression in mouse neural progenitor cells differentiated from mES and fixed overnight in 70% ethanol (Red curve). PE-labelled goat anti-mouse IgG was used as secondary antibody. Negative control processed with secondary antibody only (Blue curve). Flow cytometry data and results were generated using Orflo MoxiflowTM instrument and protocols.
Analysis by western blot of pNF-H expression in tissue lysates with M-1387-50, Neurofilament heavy polypeptide, phosphorylated (pNF-H), Clone NAP4, Mouse mAb, (green, 1:10,000) with a strong band at approx 200-220kDa corresponding to the phosphorylated form of the NF-H subunit. A minor band is observed at 160kDa corresponding to the non-phosphorylated NF-H. Smaller proteolytic fragments of NF-H are also detected in spinal cord preparations. Lane 1: MWM; Lane 2: rat brain; Lane 3: rat spinal cord; Lane 4: mouse brain; Lane 5: mouse spinal cord; Lane 6: pig spinal cord; Lane 7: cow spinal cord.
Specific ReferencesBoylan K. et al (2009) Immunoreactivity of the phosphorylated axonal neurofilament H subunit (pNF-H) in blood of ALS model rodents and ALS patients: evaluation of blood pNF-H as a potential ALS biomarker. J Neurochem. 2009 Dec;111(5):1182-91. Rangaraju S. et al (2009) Molecular architecture of myelinated peripheral nerves is supported by calorie restriction with aging. Aging Cell. 2009 Apr;8(2):178-91.
1800 605-5127
+61 (0)8 8352 7711
