Anti-Nefh Antibody Picoband™

Product Name

Anti-Nefh Antibody Picoband™

View all NF-H Antibodies

SKU/Catalog Number

A05307

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-Nefh Antibody Picoband™ catalog # A05307. Tested in ELISA, Flow Cytometry, IHC, WB applications. This antibody reacts with Mouse, Rat.

Storage & Handling

Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.

Cite This Product

Anti-Nefh Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A05307)

Host

Rabbit

Contents

Each vial contains 4mg Trehalose, 0.9mg NaCl and 0.2mg Na2HPO4.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E.coli-derived mouse Nefh recombinant protein (Position: Y109-E466).

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins.

Reactive Species

A05307 is reactive to NEFH in Mouse, Rat

Applications

A05307 is guaranteed for ELISA, Flow Cytometry, IHC, WB Boster Guarantee

Observed Molecular Weight

117-220 kDa

Calculated molecular weight

112.479kDa

Background of NF-H

Neurofilaments are type IV intermediate filament heteropolymers composed of light, medium, and heavy chains. Neurofilaments comprise the axoskeleton and functionally maintain neuronal caliber. They may also play a role in intracellular transport to axons and dendrites. This gene encodes the heavy neurofilament protein. This protein is commonly used as a biomarker of neuronal damage and susceptibility to amyotrophic lateral sclerosis (ALS) has been associated with mutations in this gene.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.1-0.25μg/ml, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Mouse, Rat
Flow Cytometry, 1-3μg/1x10⁶ cells, Mouse
Direct ELISA, 0.1-0.5μg/ml, Mouse

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of Nefh using anti-Nefh antibody (A05307).
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 rabbit anti-Nefh antigen affinity purified polyclonal antibody (Catalog # A05307) at 0.25 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-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Nefh at approximately 117-220KD. The expected band size for Nefh is at 117KD.

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Figure 2. Flow Cytometry analysis of Neuro-2a cells using anti-Nefh antibody (A05307).
Overlay histogram showing Neuro-2a cells stained with A05307 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Nefh Antibody (A05307, 1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

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Figure 3. IHC analysis of Nefh using anti-Nefh antibody (A05307).
Nefh was detected in paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-Nefh Antibody (A05307) overnight at 4°C. Biotinylated goat anti-rabbit 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 # SA1022) with DAB as the chromogen.

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Figure 4. IHC analysis of Nefh using anti-Nefh antibody (A05307).
Nefh was detected in paraffin-embedded section of mouse spinal cord tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-Nefh Antibody (A05307) overnight at 4°C. Biotinylated goat anti-rabbit 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 # SA1022) with DAB as the chromogen.

Gene/Protein Information For NEFH (Source: Uniprot.org, NCBI)

Alternative Names

200 kDa neurofilament protein; KIAA0845; NEFH; Neurofilament Heavy (200kDa); neurofilament heavy polypeptide; Neurofilament triplet H protein; neurofilament, heavy polypeptide 200kDa; neurofilament, heavy polypeptide; NF200; NFH; NF-H NEFH CMT2CC, NFH neurofilament heavy neurofilament heavy polypeptide|200 kDa neurofilament protein|NF-H|neurofilament triplet H protein|neurofilament, heavy polypeptide 200kDa

For more info on NEFH, check out the NEFH Infographic

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In this infographic, you will see the following information for NEFH: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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