Neurotrophin 4 (NTF4) ELISA Kit

Details for Product No. ABIN625066
Request Want additional data for this product?

The Independent Validation Initiative strives to provide you with high quality data. Find out more

Target Name (Antigen)
Reactivity
Alternatives Human
Kits with alternative reactivity to:
(3), (2), (6), (2), (6), (12), (5), (12), (3), (5), (10), (2)
Methode Type Sandwich ELISA
Application
ELISA
Pubmed 2 references available
Quantity 1 kit
Options
Shipping to United States (Change)
Availability Discontinued
Request Want additional data for this product?

The Independent Validation Initiative strives to provide you with high quality data. Find out more

Catalog No. ABIN625066
455.40 $
Plus shipping costs $45.00
Add to Basket

Order hotline:

  • +1 404 474 4654
  • +1 888 205 9894 (TF)
Purpose Quantitative measurement
Sample Type Serum, Plasma, Cell Culture Supernatant, Urine
Detection Method Colorimetric
Specificity This ELISA kit shows no cross-reactivity with any of the cytokines tested (e.g., human Angiogenin, BDNF, BLC, ENA-78, FGF-4, IL-1 alpha, IL-1beta, IL-2, IL-3, IL-4, IL-5, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12 p70, IL-12 p40, IL-13, IL-15, IL-309, IP-10, G-CSF, GM-CSF, IFN- gamma, Leptin, MCP-1, MCP-2, MCP-3, MDC, MIP-1 alpha, MIP-1 beta, MIP-1delta, PARC, PDGF, RANTES, SCF, TARC, TGF- beta, TIMP-1, TIMP-2, TNF-alpha, TNF-beta, TPO, VEGF).
Sensitivity The minimum detectable dose of NT-4 is typically less than 2 pg/ml.
Characteristics Human NT-4 ELISA Kit For Serum, Plasma, Cell Culture Supernatant and Urine
Components 1. NT-4 Microplate (Item A): 96 wells (12 strips x 8 wells) coated with anti-human NT-4.
2. Wash Buffer Concentrate (20x) (Item B): 25 mL of 20x concentrated solution
3. Standards (Item C): 2 vials, recombinant human NT-4.
4. Assay Diluent A (Item D): 30 mL of animal serum with 0.09 % sodium azide as preservative. For Standard/Sample (serum/plasma) diluent.
5. Assay Diluent B (Item E): 15 mL of 5x concentrated buffer. For Standard/Sample (cell culture medium/urine) diluent.
6. Detection Antibody NT-4 (Item F): 2 vial of biotinylated anti-human NT-4 (each vial is enough to assay half microplate).
7. HRP-Streptavidin Concentrate (Item G): 8 µL of 35,000x concentrated HRP-conjugated streptavidin.
8. TMB One-Step Substrate Reagent (Item H): 12 mL of 3,3™,5,5™-tetramethylbenzidine (TMB) in buffered solution.
9. Stop Solution (Item I): 8 mL of 2 M sulfuric acid.
Material not included 1. Microplate reader capable of measuring absorbance at 450 nm. 2. Precision pipettes to deliver 2 µl to 1 ml volumes. 3. Adjustable 1-25 ml pipettes for reagent preparation. 4. 100 ml and 1 liter graduated cylinders. 5. Absorbent paper. 6. Distilled or deionized water. 7. Log-log graph paper or computer and software for ELISA data analysis. 8. Tubes to prepare standard or sample dilutions.
Alternative Name NT-4
Background The Human NT-4 ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of human NT-4 in serum, plasma, cell culture supernatants and urine. This assay employs an antibody specific for human NT-4 coated on a 96-well plate. Standards and samples are pipetted into the wells and NT-4 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-human NT-4 antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of NT-4 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm. Reproducibility: Intra-Assay: CV<10% Inter-Assay: CV<12%.
Plate Pre-coated,96 wells
Reagent Preparation 1. Bring all reagents and samples to room temperature (18 - 25°C) before use. 2. Sample dilution: If your samples need to be diluted, Assay Diluent A (Item D) is used for dilution of serum/plasma samples, and Assay Diluent B (Item E) is used for dilution of culture supernatants and urine. 3. Assay Diluent B should be diluted 5-fold with deionized or distilled water. 4. Preparation of standard: Briefly spin the vial of Item C and then add 400 µl Assay Diluent A (for serum/plasma samples) or 1x Assay Diluent B (for cell culture medium and urine, Assay Diluent B should be diluted 5-fold with deionized or distilled water) into Item C vial to prepare a 50 ng/ml standard. Dissolve the powder thoroughly by a gentle mix. Add 20 µl NT-4 standard from the vial of Item C, into a tube with 980 µl Assay Diluent A or 1x Assay Diluent B to prepare a 1,000 pg/ml stock standard solution. Pipette 400 µl Assay Diluent A or 1x Assay Diluent B into each tube. Use the stock standard solution to produce a dilution series (shown below). Mix each tube thoroughly before the next transfer. Assay Diluent A or 1x Assay Diluent B serves as the zero standard (0 pg/ml). 5. If the Wash Concentrate (20x) (Item B) contains visible crystals, warm to room temperature and mix gently until dissolved. Dilute 20 ml of Wash Buffer Concentrate into deionized or distilled water to yield 400 ml of 1x Wash Buffer. 6. Briefly spin the Detection Antibody vial (Item F) before use. Add 100 µl of 1x Assay Diluent B into the vial to prepare a detection antibody concentrate. Pipette up and down to mix gently (the concentrate can be stored at 4°C for 5 days). The detection antibody concentrate should be diluted 80-fold with 1x Assay Diluent B and used in step 4 of Part VI Assay Procedure. 7. Briefly spin the HRP-Streptavidin concentrate vial (Item G) and pipette up and down to mix gently before use. HRP- Streptavidin concentrate should be diluted 35,000-fold with 1x Assay Diluent B. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 2 µl of HRP-Streptavidin concentrate into a tube with 198.0 µl 1x Assay Diluent B to prepare a 100-fold diluted HRP- Streptavidin solution (do not store the diluted solution for next day use). Mix through and then pipette 40 µl of prepared 100-fold diluted solution into a tube with 14 ml 1x Assay Diluent B to prepare a final 35,000 fold diluted HRP-Streptavidin solution.
Assay Procedure 1. Bring all reagents and samples to room temperature (18 - 25°C) before use. It is recommended that all standards and samples be run at least in duplicate. 2. Add 100 µl of each standard (see Reagent Preparation step 2) and sample into appropriate wells. Cover well and incubate for 2.5 hours at room temperature or over night at 4°C with gentle shaking. 3. Discard the solution and wash 4 times with 1x Wash Solution. Wash by filling each well with Wash Buffer (300 µl) using a multi-channel Pipette or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels. 4. Add 100 µl of 1x prepared biotinylated antibody (Reagent Preparation step 6) to each well. Incubate for 1 hour at room temperature with gentle shaking. 5. Discard the solution. Repeat the wash as in step 3. 6. Add 100 µl of prepared Streptavidin solution (see Reagent Preparation step 7) to each well. Incubate for 45 minutes at room temperature with gentle shaking. 7. Discard the solution. Repeat the wash as in step 3. 8. Add 100 µl of TMB One-Step Substrate Reagent (Item H) to each well. Incubate for 30 minutes at room temperature in the dark with gentle shaking. 9. Add 50 µl of Stop Solution (Item I) to each well. Read at 450 nm immediately.
Calculation of Results Calculate the mean absorbance for each set of duplicate standards, controls and samples, and subtract the average zero standard optical density. Plot the standard curve on log-log graph paper or using Sigma plot software, with standard concentration on the x-axis and absorbance on the y-axis. Draw the best-fit straight line through the standard points.
Restrictions For Research Use only
Storage 4 °C
Product cited in: Kalinowska-Łyszczarz, Pawlak, Michalak et al.: "Immune cell NT-3 expression is associated with brain atrophy in multiple sclerosis patients." in: Journal of neuroimmunology, 2011 (PubMed).

Browne, Yu, Huang et al.: "Proteomic identification of neurotrophins in the eutopic endometrium of women with endometriosis." in: Fertility and sterility, Vol. 98, Issue 3, pp. 713-9, 2012 (PubMed).

Validation Images
Did you look for something else?
back to top