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Human beta-NGF

Human beta-NGF DuoSet ELISA Summary

Assay   Type

Solid   Phase Sandwich ELISA

Format

96-well   strip plate

Assay   Length

4.5   hours

Sample   Type & Volume Required Per Well

100 μL

Sensitivity

10 pg/mL

Assay   Range

31.2 -   2,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma,   Citrate Plasma)

Specificity

Natural   and recombinant human CCL3/MIP-1 alpha

 

Cross-reactivity

<   0.5% cross-reactivity observed with available related molecules.< 50%   cross-species reactivity observed with species tested

Interference

No   significant interference observed with available related molecules.

Product Summary

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human beta-NGF. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

Preparation and Storage

Shipping

The   product is shipped at ambient temperature. Upon receipt, store it immediately   at the temperature recommended below.

Storage

Store   the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: beta-NGF

Nerve growth factor (NGF) was initially isolated in the mouse submandibular gland over three decades ago as a 7S complex composed of three non- covalently linked subunits, alpha, beta, and gamma. It is now known that both the alpha and gamma subunits of NGF are members of the kallikrein family of serine proteases, while the beta subunit, called beta-NGF or 2.5S NGF, exhibits all the biological activities ascribed to NGF. The human protein shares approximately 90% identity at the amino acid level with both mouse and rat beta-NGF.

Long   Name:

beta   Nerve Growth Factor

Entrez   Gene IDs:

4803   (Human); 18049 (Mouse); 310738 (Rat); 403402 (Canine); 103350089 (Rabbit)

Alternate   Names:

Beta   nerve growth factor precursor; beta-nerve growth factor; betaNGF; beta-NGF;   HSAN5; MGC161426; nerve growth factor (beta polypeptide); nerve growth   factor, beta subunit; NGFB; NGFBMGC161428

Assay Procedure

Refer to the product datasheet for the complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.

1.     Prepare all reagents, standard dilutions, and samples as directed in the product insert.

2.     Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

3.     Add 50 μL of Assay Diluent to each well.

4.     Add 50 μL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.

5.     Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

6.     Add 100 μL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.

7.     Aspirate and wash 5 times.

8.     Add 100 μL Substrate Solution to each well.

9.     Add 100 μL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

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