NUCB2 ELISA Kit

Catalog No. ABIN1706085

This Human NUCB2 ELISA Kit is a Colorimetric ELISA Kit designed to quantify Human NUCB2. This ELISA Kit has been cited in 2+ publications.

Quick Overview for NUCB2 ELISA Kit (ABIN1706085)

Target: NUCB2 (Nucleobindin 2 (NUCB2))

Binding Specificity: AA 25-106

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 31.2-2000 pg/mL

Application: ELISA

Sample Type: Cell Culture Supernatant, Serum

Product Details NUCB2 ELISA Kit

Minimum Detection Limit: 31.2 pg/mL

Purpose: Sandwich High Sensitivity ELISA kit for Quantitative Detection of Human Nesfatin-1

Brand: PicoKine™

Analytical Method: Quantitative

Specificity: Expression system for standard: E.coli
Immunogen sequence: V25-L106

Cross-Reactivity (Details): There is no detectable cross-reactivity with other relevant proteins.

Sensitivity: <10pg/mL

Material not included: 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

Immunogen: Expression system for standard: E.coli
Immunogen sequence: V25-L106

Application Details

Application Notes: Before using Kit, spin tubes and bring down all components to bottom of tube. Duplicate well assay was recommended for both standard and sample testing.

Comment:

Sequence similarities: Belongs to the nucleobindin family.

Tissue Specificity: Predominantly expressed in spleen, testis and normal stomach. .

Plate: Pre-coated

Protocol: human Nesfatin-1 ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for Nesfatin-1 has been precoated onto 96-well plates. Standards(E.coli, V25-L106) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for Nesfatin-1 is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the human Nesfatin-1 amount of sample captured in plate.

Assay Procedure:

Aliquot 0.1 mL per well of the 2000pg/mL,1000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, 31.2pg/mL human Nesfatin-1 standard solutions into the precoated 96-well plate. Add 0.1 mL of the sample diluent buffer into the control well (Zero well). Add 0.1 mL of each properly diluted sample of human cell culture supernates or serum to each empty well. See "Sample Dilution Guideline" above for details. It is recommended that each human Nesfatin-1 standard solution and each sample be measured in duplicate. Seal the plate with the cover and incubate at 37 °C for 90 min.

Assay Precision:

• Sample 1: n=16, Mean(pg/ml): 146, Standard deviation: 7.6, CV(%): 5.2
• Sample 2: n=16, Mean(pg/ml): 726, Standard deviation: 35.6, CV(%): 4.9
• Sample 3: n=16, Mean(pg/ml): 1252, Standard deviation: 68.86, CV(%): 5.5,
• Sample 1: n=24, Mean(pg/ml): 177, Standard deviation: 9.56, CV(%): 5.4
• Sample 2: n=24, Mean(pg/ml): 838, Standard deviation: 47.8, CV(%): 5.7
• Sample 3: n=24, Mean(pg/ml): 1527, Standard deviation: 96.2, CV(%): 6.3

Restrictions: For Research Use only

Handling

Handling Advice: Avoid multiple freeze-thaw cycles.

Storage: -20 °C,4 °C

Storage Comment: Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles

Expiry Date: 12 months

References for NUCB2 ELISA Kit (ABIN1706085)

Algul, Ozdenk, Ozcelik: "Variations in leptin, nesfatin-1 and irisin levels induced by aerobic exercise in young trained and untrained male subjects." in: Biology of sport, Vol. 34, Issue 4, pp. 339-344, (2018) (PubMed).

Çatl?, An?k, Küme, Çalan, Dündar, Böber, Abac?: "Serum nesfatin-1 and leptin levels in non-obese girls with premature thelarche." in: Journal of endocrinological investigation, Vol. 38, Issue 8, pp. 909-13, (2015) (PubMed).

Target Details for NUCB2

Target: NUCB2 (Nucleobindin 2 (NUCB2))

Alternative Name: NUCB2

Background:

Protein Function: Calcium-binding protein. May have a role in calcium homeostasis.

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.

Synonyms: Nucleobindin-2,DNA-binding protein NEFA,Gastric cancer antigen Zg4,Prepronesfatin,Nesfatin-1,NUCB2,NEFA,

Full Gene Name: Nucleobindin-2

Cellular Localisation: Golgi apparatus . Membrane, Peripheral membrane protein . Cytoplasm . Secreted . Endoplasmic reticulum . Nucleus envelope . Golgi retention is mediated by its N-terminal region.

Gene ID: 4925

UniProt: P80303