ELISA 2.0 kit for measuring Aspergillus fumigatus allergen, Asp f 1.
|Limit of Detection:||0.32ng/mL|
|Background:||OD<0.08 at 450nm|
|Coefficient of Determination:||R-squared>0.98|
|Contents:||Plate: Pre-coated with anti-Asp f 1 monoclonal antibody 4A6
Vial 1: (white top) nAsp f 1 allergen standard
Vial 2: (brown) Rabbit anti-Asp f 1
Vial 3: (red top) Peroxidase conjugated Goat anti. Rabbit IgG
Bottle 1: Wash buffer, (10x concentrate)
Bottle 2: Assay buffer, (10x concentrate)
Bottle 3: TMB developing substrate
Bottle 4: Stop solution (0.5N sulfuric acid)
|Storage:||Store at 2-8°C.|
|Expiry:||6 months from date of receipt|
|Not Provided:||Type I ultrapure water or 18.2MΩ de-ionized water
Volumetric measuring equipment (e.g. serological pipette, graduated cylinder)
Clean containers for buffer and reagent preparation
Calibrated single and multi-channel micropipettes and tips
Plate reader capable of reading absorbance at 450nm
Analysis software (recommended, but not required)
|Notes:||The allergen standard is recommended for immunoassay calibration
A list of frequently asked questions and troubleshooting guide can be found under the ‘Support’ tab on our website.
For research and commercial use in vitro: not for human in vivo or therapeutic use.
|Certificate of Analysis|
|Pre-coated Plate:||96-well polystyrene microtiter plate coated with
monoclonal antibody 4A6 and treated with stabilizing
agent. Sealed in foil pouch with desiccant.
|Immunogen:||Asp f 1|
|Specificity:||Binds to a common epitope on Aspergillus fumigatus allergen, Asp f 1.|
|Purification:||Produced in tissue culture and purified by chromatography using Protein G. Single heavy and light chain bands on SDS-PAGE.|
|Detection Antibody:||Polyclonal antibody raised against partially purified Asp f 1 (1,2).|
|Vial:||Brown Cap; 100µl|
|Specificity:||The pAb contains IgG antibodies to Asp f 1 as well as IgG antibodies to other Aspergillus allergens.|
|Activity:||The pAb is in phosphate buffered saline containing 1%BSA 50%glycerol PBS, pH 7.4. The pAb has been 0.22µm filtered and should be diluted 1/1000 for use in Asp f 1 ELISA.|
|Allergen Standard:||Naturally purified Asp r 1 prepared in 1% BSA/50% glycerol/PBS, 0.22μm filtered, preservative free, pH 7.4|
The Asp f 1 concentration was determined by amino acid analysis. Asp f 1 and Asp r 1 (mitogillin) are 95% homologous (differing by 4 amino acid residues) and are antigenically and functionally indistinguishable (1).
|1. Arruda, LK, Platts-Mills, TAE, Fox, JW and Chapman, MD (1990). Aspergillus fumigatus allergen I, a major IgE binding protein, is a member of the mitogillin family of cytotoxins. J. Exp. Med., 172:1529-1532.
2. Arruda LK, Platts-Mills TAE, Longbottom JL, Chapman MD (1992). Aspergillus fumigatus: Identification of 16kd, 18kd and 45kd antigens recognized by human IgG and IgE antibodies and murine monoclonal antibodies. J. Allergy Clin. Immunol. 89:1166-1176.
3. Arruda LK, Mann BJ, Chapman MD (1992). Selective expression of a major allergen and cytotoxin, Asp f 1, in Aspergillus fumigatus. J. Immunol. 149:3354- 3359.
4. Sporik RB, Arruda LK, Woodfolk J, Chapman MD, Platts-Mills TAE (1993). Environmental exposure to Aspergillus fumigatus allergen 1 (Asp f 1). Clin. Exp. Allergy 23:326-331.
5. El-Dahr JM, Fink R, Selden R, Arruda LK, Platts-Mills TAE, Heymann PW (1994). Development of immune responses to Aspergillus at an early age in children with cystic fibrosis. Am. J.
Respir. Crit. Care Med. 150:1513-1518.
6. Vailes LD, Sridhara S, Cromwell O, Weber B, Breitenbach M, Chapman MD (2001). Quantitation of the major fungal allergens, Alt a 1 and Asp f 1, in commercial allergenic products. J. Allergy Clin. Immunol. 107:641-646.
7. Chapman MD (2006). Challenges associated with indoor moulds: Health effects, immune response and exposure assessment. Medical Mycology. 44:S29-S32.
|(For complete reference list see EPC-AF1 Certificate of Analysis)|