|Validated Species:||Human, Mouse, Rat|
|Application:||IHC-P, IHC-F, WB|
Data & Images
|Product Name||Anti-NEFH Antibody (Monoclonal, N52)|
|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) N52 (Boster Biological Technology, Pleasanton CA, USA, Catalog # MA1071)|
|Replacement Item||This antibody may replace the following items: sc-133165|sc-133237|sc-137009|sc-20014|sc-20015|sc-20112|sc-22909|sc-32273|sc-32728|sc-32729|sc-32730|sc-51682|sc-52350|sc-53798|sc-56575|sc-58553 from Santa Cruz Biotechnology.|
|Validated Species||Human, Mouse, Rat|
|Application||IHC-P, IHC-F, WB
*Our Boster Guarantee covers the use of this product in the above tested applications.
**For positive and negative control design, consult "Tissue specificity" under Protein Target Info.
|Recommended Detection Systems||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).
*Blocking peptide can be purchased at $50. Contact us for more information
**Boster also offers various secondary antibodies for Immunoflourescecne and IHC. Take advantage of the buy 1 primary antibody get 1 secondary antibody for free promotion for the entire year 2018!
|Immunogen||C-terminal segment of enzymatically dephosphorylated pig Neurofilament 200.|
|Cross Reactivity||No cross reactivity with other proteins|
|Contents||Mouse ascites fluid, 1.2% sodium acetate, 2mg BSA, with 0.01mg NaN3 as preservative.|
|Concentration||Add 1ml of PBS buffer will yield a concentration of 100ug/ml.|
|Storage||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.|
Protein Target Info (Source: Uniprot.org)
You can check the tissue specificity below for information on selecting positive and negative control.
|Protein Name||Neurofilament heavy polypeptide|
|Molecular Weight||115378 MW|
|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.|
|Sequence Similarities||Belongs to the intermediate filament family.|
|Alternative Names||Neurofilament heavy polypeptide;NF-H;200 kDa neurofilament protein;Neurofilament triplet H protein;Nefh;Nfh;|
|Research Areas|||neuroscience|neurology process|neurodegenerative disease|alzheimer's disease| neuroscience|cell adhesion proteins|cytoskeletal proteins|intermediate filaments| signal transduction|cytoskeleton / ecm|cytoskeleton|class iv|neurofilaments|cell type marker|neuron marker|axon marker||
Background for Neurofilament heavy polypeptide
Anti-NEFH Antibody (Monoclonal, N52) Images
Click the images to enlarge.
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.
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.