( immunoglobulins) are gamma globulin proteins that are found in blood and are used by the immune system to identify and neutralize foreign objects
The
antibody recognises and bind to the
antigenic determinant region of the antigen
Specific
antibodies are produced by
injecting an antigen into a mammal, such as a mouse.
Definitions- cont
Antigens Suatu bahan biokimia yang berada di dalam tubuh akan menstimulasi produksi antibody Is present naturally in the body like hormones Is not present in the body in normal condition like drugs
Immunoassay Teknik pengujian yang dilakukan dengan menggunakan ikatan antara antigen dan antibody yang sesuai agar teridentifikasi kuantitas specific antigen or antibody in a sample
Definitions- cont Analyte
Sampel yang dianalisis menggunakan
immunoasssays. Analit bisa berupa Antibody or Antigen
ELISA technique Yaitu cara pengujian bioassay menggunakan uji immunology untuk mendeteksi keberadaan antibody or an antigen in a sample. The
technique is divided into
1- Competitive ELISA 2- Sandwich ELISA (also called direct ELISA) Indirect ELISA- 3 Non Competitive-4
Competitive ELISA
Antigen berlabel dan antigen sampel berkompetisi
mengikat antibodi. Banyaknya antigen sampel yang terikat antibodi diukur dari pembentukan warna
:Tahapan Uji Antibodi dilapiskan pada mikroplat Penambahan hapten-enzim konjugat dan larutan contoh Terjadi kompetisi analit contoh dan enzim untuk mengikat antibodi Penambahan Substrat pembentuk warna
KOMPETISI LANGSUNG (SANDWICH)
Analit sampel (antigen) ditempelkan pada plat antibodi. Setelah pencucian matriks sampel, antigen sampel ditempelkan antigen berlabel). Banyaknya antigen sampel yang dapat mengikat antibodi ditentukan dengan penambahan pereaksi pembentuk warna
Sandwich ELISA
Indirect ELISA Analit sampel dan antigen berlabel tidak langsung berkompetisi mengikat antibodi, banyaknya analit yang berikatan dengan antibodi, menunjukkan banyaknya antigen berlabel. Pada metode ini yang terukur adalah jumlah antigen berlabel yang terwarnai oleh .pereaksi
Indirect ELISA
An example of an ELISA experiment
Before
starting the work read kit
instruction carefully
1-
The 96 well plate is labeled
carefully and the first wells are used to draw the standard curve
An example of an ELISA experiment-Cont
The
sample is added to plate in
duplicate or triplicate and then the mean result is calculated
The
quality control sample which is
provided with the kit is treated as the test samples
Results After
reading the results the standard
curve is drawn were the concentration is blotted on the X-axis and the absorbance on the Y-axis
Absorptio n nm
Concentration ng/ml
Results-cont The
standards concentrations is specified on the x-axis and the reading of each standard is specified on the y-axis and the standard curve is drawn
