Anti-NEFH Antibody (Monoclonal, N52)
|Reactivity||Human, Mouse, Rat|
|Ig Isotype||Mouse IgG1|
|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)|
|Contents/Buffer||Mouse ascites fluid, 1.2% sodium acetate, 2mg BSA, with 0.01mg NaN3 as preservative.|
|Immunogen||C-terminal segment of enzymatically dephosphorylated pig Neurofilament 200.|
|Reactivity||Human, Mouse, Rat|
Assay Dilutions Overview
Immunohistochemistry(Paraffin-embedded Section), 1-2μg/ml, Human, mouse, rat, By Heat
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 1Î¼g/ml mouse anti- NEFH Antibody (MA1071) overnight at 4Â°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37Â°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 Î¼g/mL overnight at 4Â°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.|
|Research Areas||Human, Mouse, Rat
*You can search these to find other products in these research areas.
|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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Q: Is your western blot protocol provided from the website applicable for all your antibodies? Keyword: applications, WBA: The protocol is applicable for all our antibodies in WB, the NC Membrane(0.45μm or 0.22μm) and transfer time(70 mins or 50 mins) depends on the protein molecular weight, details can be found in included protocol.
Q: Can I conjugate markers to this antibody? Can I link custom conjugates to this antibody? Keyword: conjugationA: The antibody is stored with BSA and cannot be conjugated with markers. Carrier free antibodies are available upon request. Please contact firstname.lastname@example.org
Q: Can you provide monoclonal antibodies carrier free?A: For most of our monoclonal antibodies, we are unable to provide them carrier free. If you would like to check if a specific antibody can be provided carrier free, please send an email to email@example.com.
Q: What are some alternative names that could be used to describe this product?A: Some common names include but are not limited to nf200 antibody, nf-h antibody