NAMPT ELISA Kit

Catalog No. ABIN6973971

Product Details NAMPT ELISA Kit

Target: NAMPT (Nicotinamide phosphoribosyltransferase (NAMPT))

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 0.625 ng/mL - 40 ng/mL

Minimum Detection Limit: 0.625 ng/mL

Application: ELISA

Purpose: For the quantitative determination of human visfatin concentrations in serum, plasma, urine, tissue homogenates.

Sample Type: Plasma, Serum, Tissue Homogenate, Urine

Analytical Method: Quantitative

Specificity: This assay has high sensitivity and excellent specificity for detection of human visfatin. No significant cross-reactivity or interference between human visfatin and analogues was observed. Note: Limited by current skills and knowledge, it is impossible for us to complete the cross-reactivity detection between human visfatin and all the analogues, therefore, cross reaction may still exist.

Sensitivity: 0.156 ng/mL

Components:

• Assay plate
• Standard
• HRP-avidin (100 x concentrate)
• Biotin-antibody (100 x concentrate)
• Sample Diluent
• HRP-avidin Diluent
• Biotin-antibody Diluent
• Wash Buffer (25 x concentrate)
• TMB Substrate
• Stop Solution
• Adhesive Strip

Application Details

Application Notes: Optimal working dilution should be determined by the investigator.

Sample Volume: 100 μL

Assay Time: 1 - 4.5 h

Plate: Pre-coated

Protocol:

1. Prepare reagents, samples and standards as instructed.
2. Add 100 µL standard or sample to each well. Incubate 2 hours at 37 °C.
3. Remove the liquid of each well, don't wash.
4. Add 100 µL Biotin-antibody (1x) to each well. Incubate 1 hour at 37 °C.
5. Aspirate and wash 3 times.
6. Add 100 µL HRP-avidin (1x) to each well. Incubate 1 hour at 37 °C
7. Aspirate and wash 5 times.
8. Add 90 μL of TMB Substrate to each well. Incubate for 15-30 minutes at 37 °C. Protect from light.
9. Add 50 µL Stop Solution to each well. Read at 450 nm within 5 minutes.

Reagent Preparation:

1. Biotin-antibody (1x) - Centrifuge the vial before opening. Biotin-antibody requires a 100-fold dilution. A suggested 100-fold dilution is 10 μL of Biotin-antibody + 990 μL of Biotin-antibody Diluent.
2. HRP-avidin (1x) - Centrifuge the vial before opening. HRP-avidin requires a 100-fold dilution. A suggested 100-fold dilution is 10 μL of HRP-avidin + 990 μL of HRP-avidin Diluent.
3. Wash Buffer (1x) - If crystals have formed in the concentrate, warm up to room temperature and mix gently until the crystals have completely dissolved. Dilute 20 mL of Wash Buffer Concentrate (25 x) into deionized or distilled water to prepare 500 mL of Wash Buffer (1 x).
4. Standard Centrifuge the standard vial at 6000-10000rpm for 30s. Reconstitute the Standard with 1.0 mL of Sample Diluent. Do not substitute other diluents. This reconstitution produces a stock solution of 40 ng/mL. Mix the standard to ensure complete reconstitution and allow the standard to sit for a minimum of 15 minutes with gentle agitation prior to making dilutions. Pipette 250 μL of Sample Diluent into each tube (S0-S6). Use the stock solution to produce a 2-fold dilution series (below). Mix each tube thoroughly before the next transfer. The undiluted Standard serves as the high standard (40 ng/mL). Sample Diluent serves as the zero standard (0 ng/mL).

Note:

• Kindly use graduated containers to prepare the reagent. Please don't prepare the reagent directly in the Diluent vials provided in the kit.
• Bring all reagents to room temperature (18-25 °C) before use for 30 min.
• Prepare fresh standard for each assay. Use within 4 hours and discard after use.
• Making serial dilution in the wells directly is not permitted.
• Please carefully reconstitute Standards according to the instruction, and avoid foaming and mix gently until the crystals have completely dissolved. To minimize imprecision caused by pipetting, use small volumes and ensure that pipettors are calibrated. It is recommended to suck more than 10 μL for once pipetting.
• Distilled water is recommended to be used to make the preparation for reagents. Contaminated water or container for reagent preparation will influence the detection result.

Sample Preparation:

• It is recommended to use fresh samples without long storage, otherwise protein degradation and denaturationmay occur in these samples, leading to false results. Samples should therefore be stored for a short periodat 2 - 8 °C or aliquoted at -20 °C (≤1 month) or -80 °C (≤ 3 months). Repeated freeze-thawcycles should be avoided. Prior to assay, the frozen samples should be slowly thawed and centrifuged toremove precipitates.
• If the sample type is not specified in the instructions, a preliminary test is necessary to determinecompatibility with the kit.
• If a lysis buffer is used to prepare tissue homogenates or cell culture supernatant, there is a possibilityof causing a deviation due to the introduced chemical substance.The recommended dilution factor is for reference only.
• Please estimate the concentration of the samples before performing the test. If the values are not in therange of the standard curve, the optimal sample dilution for the particular experiment has to be determined.Samples should then be diluted with PBS (pH =7.0-7.2).

Assay Precision: Intra-assay Precision (Precision within an assay): CV%<8% Three samples of known concentration were tested twenty times on one plate to assess.
Inter-assay Precision (Precision between assays): CV%<10% Three samples of known concentration were tested in twenty assays to assess.

Restrictions: For Research Use only

Handling

Storage: 4 °C,-20 °C

Storage Comment: Unopened kit Store at 2 - 8°C. Do not use the kit beyond the expiration date May be stored for up to 1 month at 2 - 8°C. Coated assay Try to keep it in a sealed aluminum foil bag, plate and avoid the damp. Standard May be stored for up to 1 month at 2 - 8° C. If Biotin-antibody don't make recent use, better keep it store at HRP-avidin -20°C. Biotin-antibody Diluent Opened kit HRP-avidin Diluent Sample May be stored for up to 1 month at 2 - 8°C. Diluent Wash Buffer TMB Substrate Stop Solution *Provided this is within the expiration date of the kit.

Expiry Date: 6 months

References for NAMPT ELISA Kit (ABIN6973971)

Behboudi-Gandevani, Ramezani Tehrani, Bidhendi Yarandi, Noroozzadeh, Hedayati, Azizi: "The association between polycystic ovary syndrome, obesity, and the serum concentration of adipokines." in: Journal of endocrinological investigation, Vol. 40, Issue 8, pp. 859-866, (2018) (PubMed).

El Shemi, Mohamed, AbdelRahman, Abdel Al, Ramadan: "Effect of intrauterine growth pattern on serum visfatin concentrations in full-term infants at birth and at 6 months of life." in: Journal of neonatal-perinatal medicine, Vol. 9, Issue 1, pp. 73-82, (2017) (PubMed).

Toru, Ayada, Genç, ?ahin, Ar?k, Acat, Bulut, Çetinkaya: "Visfatin and ghrelin: can they be forthcoming biomarkers or new drug targets for asthma?" in: International journal of clinical and experimental medicine, Vol. 8, Issue 4, pp. 6257-61, (2015) (PubMed).

Nääv, Erlandsson, Axelsson, Larsson, Johansson, Wester-Rosenlöf, Mörgelin, Casslén, Gram, Åkerström, Hansson: "A1M Ameliorates Preeclampsia-Like Symptoms in Placenta and Kidney Induced by Cell-Free Fetal Hemoglobin in Rabbit." in: PLoS ONE, Vol. 10, Issue 5, pp. e0125499, (2015) (PubMed).

Castro-Piñero, Carbonell-Baeza, Martinez-Gomez, Gómez-Martínez, Cabanas-Sánchez, Santiago, Veses, Bandrés, Gonzalez-Galo, Gomez-Gallego, Veiga, Ruiz, Marcos et al.: "Follow-up in healthy schoolchildren and in adolescents with Down syndrome: psycho-environmental and genetic determinants of physical activity and its impact on fitness, cardiovascular diseases, ..." in: BMC public health, Vol. 14, pp. 400, (2014) (PubMed).

McMillin, Frampton, Thompson, Galindo, Standeford, Whittington, Alpini, DeMorrow: "Neuronal CCL2 is upregulated during hepatic encephalopathy and contributes to microglia activation and neurological decline." in: Journal of neuroinflammation, Vol. 11, pp. 121, (2014) (PubMed).

Rafraf, Mohammadi, Asghari-Jafarabadi, Farzadi: "Omega-3 fatty acids improve glucose metabolism without effects on obesity values and serum visfatin levels in women with polycystic ovary syndrome." in: Journal of the American College of Nutrition, Vol. 31, Issue 5, pp. 361-8, (2013) (PubMed).

Target Details for NAMPT

Target: NAMPT (Nicotinamide phosphoribosyltransferase (NAMPT))

Alternative Name: nicotinamide phosphoribosyltransferase (NAMPT Products)

Synonyms: 1110035O14Rik ELISA Kit, PBEF ELISA Kit, PBEF1 ELISA Kit, VF ELISA Kit, VISFATIN ELISA Kit, AI314458 ELISA Kit, AI480535 ELISA Kit, NAmPRTase ELISA Kit, Pbef ELISA Kit, Pbef1 ELISA Kit, Visfatin ELISA Kit, visfatin ELISA Kit, pbef ELISA Kit, pbef1 ELISA Kit, nicotinamide phosphoribosyltransferase ELISA Kit, Nicotinamide phosphoribosyltransferase ELISA Kit, nicotinamide phosphoribosyltransferase S homeolog ELISA Kit, NAMPT ELISA Kit, Nampt ELISA Kit, nampt ELISA Kit, Smon_0680 ELISA Kit, Mrub_2845 ELISA Kit, Cseg_3704 ELISA Kit, BC1002_3997 ELISA Kit, Ftrac_2686 ELISA Kit, Varpa_5681 ELISA Kit, Celal_3900 ELISA Kit, Deima_0880 ELISA Kit, BC1001_3724 ELISA Kit, Celly_1696 ELISA Kit, Clole_1542 ELISA Kit, Tsp_05934 ELISA Kit, nampt.S ELISA Kit

Background:

Abbreviation: NAMPT

Alias: 1110035O14Rik, DKFZp666B131, MGC117256, PBEF, PBEF1, VF, VISFATIN, NAmPRTase|pre-B cell-enhancing factor|pre-B-cell colony enhancing factor 1

UniProt: P43490