|Product Name||Human Nesfatin-1 PicoKine™ ELISA Kit|
|Storage & Handling||Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles(Shipped with wet ice.)|
|Size||96wells/kit, with removable strips.|
|Description||Sandwich High Sensitivity ELISA kit for Quantitative Detection of Human Nesfatin-1. 96wells/kit, with removable strips.|
|Cite This Product||Human Nesfatin-1 PicoKine™ ELISA Kit (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK1138)|
|Sample Type||cell culture supernates and serum..|
|Immunogen||Expression system for standard: E.coli; Immunogen sequence: V25-L106|
|Cross Reactivity||There is no detectable cross-reactivity with other relevant proteins.|
|Antibody Clonalities||Capture Antibody | Detection Antibody:
monoclonal antibody from mouse|polyclonal antibody from goat
|EK1138-CAP||96-well plate precoated with anti-Human NUCB2 antibody||1|
|EK1138-ST||lyophilized recombinant Human NUCB2 standard||2ng/tube|
|EK1138-DA||biotinylated anti-Human NUCB2 antibody||130ul|
|AR1106-1||sample diluent buffer||30ml|
|AR1106-2||antibody diluent buffer||12ml|
|AR1106-3||ABC diluent buffer||12ml|
|AR1104||TMB color developing agent||10ml|
|AR1105||TMB stop solution||10ml|
*Why there is no wash buffer? Our Avidin-Biotin-Peroxidase Diluent contains the detergent (TWEEN) normally present in other companies' ELISA kits. This saves you the step of having to wash with the special wash buffer and achieve similar or better signal to noise ratio. The wash can use regular wash buffers (PBS, TBS etc.) commonly found in labs.
Materials Required But Not Provided
- Microplate reader in standard size.
- Automated plate washer.
- Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection.
- Clean tubes and Eppendorf tubes.
- Washing buffer (neutral PBS or TBS).
- Preparation of 0.01M TBS: Add 1.2g Tris, 8.5g NaCl; 450μl of purified acetic acid or 700μl of concentrated hydrochloric acid to 1000ml H2
- Preparation of 0.01 M PBS: Add 8.5g sodium chloride, 1.4g Na O and adjust pH to 7.2-7.
Typical Data Obtained from Human Nesfatin-1 PicoKine™ ELISA Kit
(TMB reaction incubate at 37°C for 15-25min)
Intra/Inter Assay Precision
|Intra-Assay Precision||Inter-Assay Precision|
Three samples with differing target protein concentrations were assayed using four different lots to measure the CV% lot to lot variance.
To assay reproducibility, three samples with differing target protein concentrations were assayed using four different lots.
|Lots||Lot1 (pg/ml)||Lot2 (pg/ml)||Lot3 (pg/ml)||Lot4 (pg/ml)||Mean (pg/ml)||Standard Deviation||CV (%)|
*The typical data is obtained from Boster's internal QC result and for reference only. It may differ from the lab test results of the end users. It is more important that the user's lab test results reflect the same linearity demonstrated in the typical data than achieving exactly the same O.D. values.
Protein Target Info (Source: Uniprot.org)
|Tissue Specificity||Predominantly expressed in spleen, testis and normal stomach. .|
|Alternative Names||Nucleobindin-2;DNA-binding protein NEFA;Gastric cancer antigen Zg4;Prepronesfatin;Nesfatin-1;NUCB2;NEFA;|
|Subcellular Localization||Golgi apparatus . Membrane ; Peripheral membrane protein . Cytoplasm . Secreted . Endoplasmic reticulum . Nucleus envelope . Golgi retention is mediated by its N-terminal region.|
|Molecular Weight||50196 MW|
*if product is indicated to react with multiple species, protein info is based on the human gene.
|Protein Function||Calcium-binding protein. May have a role in calcium homeostasis.|
|Research Areas||Calcium Binding Proteins, Calcium Signaling, Chromatin Binding Proteins, Dna / Rna Binding, Epigenetics And Nuclear Signaling, Signal Transduction, Signaling Pathway
*You can search these to find other products in these research areas.
|Background||Nesfatin-1 is a naturally occurring protein molecule produced by the brains of mammals. It is responsible for regulating appetite and production of body fat. Nesfatin-1 is a metabolic polypeptide encoded in the N-terminal region of the protein precursor, Nucleobindin2(NUCB2). Recombinant human Nesfatin-1 is a 9.7 kDa protein containing 82 amino acid residues. Originally identified as a hypothalamic neuropeptide, Nesfatin-1 is also expressed in other areas of the brain, and in pancreatic islets of Langerhans, gastric endocrine cells and adipocytes. Nesfatin-1 plays a role in hunger and energy regulation in an independent manner.|
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1. Diluent the samples with the provided sample diluent buffer into 100ul.
2. Add 50ul of standard solution, when 50ul of sample will be added into a well.
• Add tissue homogenates into the wells and then add ABC and TMB without adding any biotinylated detection antibody to see if any signal will be observed.
• If no signal is produced, then you can work on the tissue sample by using the kit.
The 1XTBS can be used if the pH value falls in the range of 7.2-7.6.
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