|Product Name||Anti-NEFH Antibody (Monoclonal, N52)|
|Storage & Handling||At -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time.Avoid repeated freezing and thawing.|
|Description||Mouse IgG monoclonal antibody for NEFH, neurofilament, heavy polypeptide (NEFH) detection. Tested with WB, IHC-P, IHC-F in Human;mouse;rat. No cross reactivity with other proteins.|
|Cite This Product||Anti-NEFH Antibody (Monoclonal, N52) (Boster Biological Technology, Pleasanton CA, USA, Catalog # MA1071)|
|Immunogen||C-terminal segment of enzymatically dephosphorylated pig Neurofilament 200.|
|Reactivity||Human, Mouse, Rat|
Assay Dilutions Overview
Immunohistochemistry(Frozen Section), 1-2μg/ml, Human, mouse, rat, -
Western blot, 0.5ml, Human, mouse, rat
Boster's Secondary Antibodies And IHC, WB Kits
The following reagents are used to generate the images below.Boster recommends Enhanced Chemiluminescent Kit with anti-Mouse IgG (EK1001) for Western blot, and HRP Conjugated anti-Mouse IgG Super Vision Assay Kit (SV0001-1) for IHC(P) and IHC(F).
Images And Assay Conditions
Figure 1. IHC analysis of NEFH using anti- NEFH antibody (MA1071).
NEFH was detected in paraffin-embedded section of rat brain tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti- NEFH Antibody (MA1071) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1021) with DAB as the chromogen.
Figure 2. Western blot analysis of NEFH using anti-NEFH antibody (MA1071).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: rat brain tissue lysates,
Lane 2: mouse brain tissue lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-NEFH antigen affinity purified monoclonal antibody (Catalog # MA1071)) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for NEFH at approximately 200KD. The expected band size for NEFH is at 112KD.
Protein Target Info (Source: Uniprot.org)
|Protein Name||Neurofilament heavy polypeptide|
|Alternative Names||Neurofilament heavy polypeptide;NF-H;200 kDa neurofilament protein;Neurofilament triplet H protein;Nefh;Nfh;|
|Molecular Weight||115378 MW|
*if product is indicated to react with multiple species, protein info is based on the human gene.
|Protein Function||Neurofilaments usually contain three intermediate filament proteins: L, M, and H which are involved in the maintenance of neuronal caliber. NF-H has an important function in mature axons that is not subserved by the two smaller NF proteins.|
|Background||Neurofilaments are composed of 3 neuron-specific proteins with apparent molecular masses of 68 kD(NFL), 125 kD(NFM), and 200 kD(NFH) on SDS-gel electrophoresis. Genomic clones for the largest human neurofilament protein(NF-H) were isolated, the intron/exon boundaries mapped and the entire protein-coding regions(exons) sequenced. mutations in neurofilaments have been linked to some forms of Charcot-Marie-Tooth disease(CMT).|
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