Calostro De La Leche

CALOKIT-VACA

INTRODUCTION AND OBJECTIVE
Colostrum is a secretion of the udder produced during first days after calving. Its mission is to confer immunitary protection for the cattle calves during the first months of their life. Colostrum composition is very different from that of raw milk. Main differences are their protein composition. While 70-80% of proteins in raw milk are caseins, in thefirst colostrum, caseins are only the 20%. Besides, whey protein concentration, responsible for some technological problems, rise from 50 to 120 times with regards to its usual level. Colostrum presence in milk can damage gravely production and standardization of dairy products. For instance, colostrum presents high concentration of some proteins with antimicrobial properties. Thus, colostrum canprevent bacterial growth responsible of cheese and milk fermentation, reducing acidification rate and altering organoleptical properties of such products. Besides, colostrum presence in milk generates problems in the cleaning and maintenance of technologic equipment, such as heat transfer, driers and pasteurizers, because of the increase in the soluble proteins ratio. Soluble proteins aredenatured easily precipitating on the surfaces of the technological equipment.

KIT COMPONENTS

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1 x Microtiter plate for 96 tests (12 strips of 8 wells) 4 x IgG standards (A,B,C,D) 1 x Conjugate Stock Solution x 20 . 1 x Substrate/Chromogen Solution (TMB). 1 x Stop Solution (Sulfuric acid 1M) Corrosive! 1 x Conjugate dilution solution.

ADDITIONAL MATERIAL

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MicropipettesPhotometer for ELISA assays

STABILITY AND STORAGE
Kit components should be stored at 4-7 ºC in darkness. Kit keeps stability for 6 months if is stored correctly.

SAFETY
Good laboratory practice should be employed when using this kit. Safety clothing should be worn and skin contact with reagents avoided. Do not ingest. A SAFETY DATA SHEET is available by request from your distributor.

7ZEU-INMUNOTEC

ENGLISH

CALOKIT-VACA

PRINCIPLE OF THE ASSAY
CALOKIT is a non competitive ELISA assay (sandwich), based on the immunoglobulins detection by means of specific antibodies. CALOKIT has been designed for quantitation of these soluble proteins (IgGs) whose concentration in colostrum is 50-100 times higher than milk. With this procedure it is possible to detect the colostrumadded from an animal to the milk from a herd of at least 50 animals. CALOKIT can quantify with precision the concentration of IgG in milk. Criteria for determining colostrum presence in milk is established by legislation in some countries, but can be considered as a general criteria that milk with a IgG concentration higher than 1-2 g/l has been adulterated with colostrum. This test can also be usedin the management of the dairy farms. Quantitation of immunoglobulins is an objective criteria to difference milk which is driven to tanks from milk that should be separated forthe calves feeding.

PREPARATION OF SAMPLES AND REAGENTS Preparation of standards:
Dilute the standards (A, B, C, D) 1/100 with distilled water. Mix 1 volume of each standard with 99 volumes of distilled water. Forinstance: 0.1 ml of standard with 9,9 ml of distilled water. Important (for a good interpretation of the result): Standards must be always used with this dilution factor . It is advisable to include a negative control of distilled water as “”zero standard” in the calibration curve.

Preparation of milk samples (final dilution 1/2000):
The samples dilution is done in two steps. An initial dilution1/100 with distilled water. For example, mixing 0,1 ml of milk and 9,9 ml of water. Then, it must be realized a second dilution 1/20 in distilled water. For example, 0,050 ml of initial solution and 0,950 ml of distilled water.

Preparation of conjugate solution:
Prepare only the solution volume needed for each assay. Mix 1 volume of conjugate stock solution with 19 volumes of conjugate dilution...