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Product Info Summary
| SKU: | A00585 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | ELISA, IHC, WB |
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Product info
Product Name
Anti-NSF Antibody Picoband®
SKU/Catalog Number
A00585
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-NSF Antibody Picoband® catalog # A00585. Tested in ELISA, IHC, WB applications. This antibody reacts with Human, Mouse, Rat. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
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-NSF Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00585)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E. coli-derived human NSF recombinant protein (Position: N620-D744).
*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
A00585 is reactive to NSF in Human, Mouse, Rat
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Observed Molecular Weight
82 kDa
Calculated molecular weight
82.594kDa
Background of NSF
N-ethylmaleimide-sensitive factor, also known as NSF, is an enzyme which in humans is encoded by the NSF gene. NSF is a homohexameric AAA ATPase involved in membrane fusion. NSF is ubiquitously found in the cytoplasm of eukaryotic cells. It is a central component of the cellular machinery in the transfer of membrane vesicles from one membrane compartment to another. During this process, SNARE proteins on two joining membranes (usually a vesicle and a target membrane such as the plasma membrane) form a tight complex. This aids fusion of the vesicle with the target membrane. It has been proposed that the role of NSF is to undo these SNARE complexes once membrane fusion has occurred, using the hydrolysis of ATP as an energy source, allowing the dissociated SNAREs to be recycled for reuse in further rounds of membrane fusion. This proposal remains controversial, however. Recent work indicates that the ATPase function of NSF does not function in recycling of vesicles but rather functions in the act of fusing vesicles with the plasma membrane.
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.
Application & Images
Applications
A00585 is guaranteed for ELISA, IHC, WB Boster Guarantee
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, 0.1-0.5μg/ml
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml
ELISA, 0.1-0.5μg/ml
Positive Control
WB: rat brain tissue, mouse brain tissue, human HepG2 cell
IHC: mouse testis tissue, rat testis tissue, human glioma tissue, human intestinal cancer tissue, human mammary cancer tissue
Validation Images & Assay Conditions
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Figure 1. Western blot analysis of NSF using anti-NSF antibody (A00585).
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,
Lane 3: human HepG2 cell 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-NSF antigen affinity purified polyclonal antibody (Catalog # A00585) 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-rabbit 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 # EK1002) with Tanon 5200 system. A specific band was detected for NSF at approximately 82KD. The expected band size for NSF is at 82KD.
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Figure 2. IHC analysis of NSF using anti-NSF antibody (A00585).
NSF was detected in paraffin-embedded section of mouse testis tissue. 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 rabbit anti-NSF Antibody (A00585) 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 3. IHC analysis of NSF using anti-NSF antibody (A00585).
NSF was detected in paraffin-embedded section of rat testis tissue. 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 rabbit anti-NSF Antibody (A00585) 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.
Click image to see more details
Figure 4. IHC analysis of NSF using anti-NSF antibody (A00585).
NSF was detected in paraffin-embedded section of human glioma tissue. 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 rabbit anti-NSF Antibody (A00585) 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.
Click image to see more details
Figure 5. IHC analysis of NSF using anti-NSF antibody (A00585).
NSF was detected in paraffin-embedded section of human intestinal cancer tissue. 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 rabbit anti-NSF Antibody (A00585) 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.
Click image to see more details
Figure 6. IHC analysis of NSF using anti-NSF antibody (A00585).
NSF was detected in paraffin-embedded section of human mammary cancer tissue. 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 rabbit anti-NSF Antibody (A00585) 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.
Protein Target Info & Infographic
Gene/Protein Information For NSF (Source: Uniprot.org, NCBI)
Gene Name
NSF
Full Name
Vesicle-fusing ATPase
Weight
82.594kDa
Superfamily
AAA ATPase family
Alternative Names
Vesicle-fusing ATPase; N-ethylmaleimide-sensitive fusion protein; NEM-sensitive fusion protein; Vesicular-fusion protein NSF; NSF NSF SEC18, SKD2 N-ethylmaleimide sensitive factor, vesicle fusing ATPase vesicle-fusing ATPase|N-ethylmaleimide-sensitive factor-like protein|N-ethylmaleimide-sensitive fusion protein|NEM-sensitive fusion protein|epididymis secretory sperm binding protein|vesicular-fusion protein NSF
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on NSF, check out the NSF Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for NSF: 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].
Specific Publications For Anti-NSF Antibody Picoband® (A00585)
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Customer Q&As
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1 Customer Q&As for Anti-NSF Antibody Picoband®
Question
We are currently using anti-NSF antibody A00585 for mouse tissue, and we are satisfied with the ELISA results. The species of reactivity given in the datasheet says human, mouse, rat. Is it true that the antibody can work on bovine tissues as well?
D. Parker
Verified customer
Asked: 2013-10-21
Answer
The anti-NSF antibody (A00585) has not been validated for cross reactivity specifically with bovine tissues, though there is a good chance of cross reactivity. We have an innovator award program that if you test this antibody and show it works in bovine you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2013-10-21
