The sections were then subjected to incubation with (LSAB
Kit/HRP,Rb/Mo/Goat (DAB+) system from DAKO cat#K0679, secondary antibodies for 15 mins and washed with PBS again.
After the reaction LSAB
+ System HRP, Dako, Poland), the antigen-ntibody complex was visualized by using DAB chromogen (S3000, DAKO, Poland).
Five genotyping methods were utilized in the current meta-analysis, including labeled streptavidin-biotin method (LSAB
), EnVision, MaxVision, avidin-biotin complex (ABC), and streptavidin-perosidase (SP) methods.
The slides were subsequently rinsed with TBS plus Tween 20, treated for 30 min at room temperature with Histofine Simple Stain rat MAX PO (M) (Nichirei, Tokyo, Japan) or a staining system kit DAKO LSAB
2 kit/HRP (DAKO Japan, Kyoto, Japan), rinsed with TBS plus Tween 20, incubated in diaminobenzidine solution containing 0.01% hydrogen peroxide for the peroxidase coloring reaction, and counterstained with Mayer's hematoxylin.
Immunostaining was performed using the labeled streptavidin biotin (LSAB
Bound antibodies were visualized using the LSAB
secondary antibody streptavidin biotin peroxidase system (DAKO).
Todas as seccoes foram submetidas a recuperacao antigenica com TRIS-EDTA (pH 9,0) em microondas por 10 minutos, incubadas com os anticorpos primarios por uma hora a 37[degrees]C e, posteriormente, com o anticorpo secundario biotinilado e o complexo LSAB
. A imunorreatividade foi observada por meio do 2-2' diaminobenzidina (DAB, Sigma-Aldrich) e a contra-coloracao utilizada foi a de hematoxilina Harris, utilizando controles positivos e negativos para cada anticorpo.
Quantification of TNF-[alpha] and IL-6 producing cells in the tonsils with TH and RT was performed on 5 pm thick sections of the tonsils, which were stained by immunohistochemical method LSAB
+ HRP (LabelledStreptAvidin-Biotin, Horst Readish Peroxidase).
Para o diagnostico e utilizado o metodo streptavidina-biotina peroxidase marcada (LSAB
) com anticorpo primario mpoliclonal anti-Pythium insidiosum na diluicao de 1:1000 em tampao fosfato salino (Phosphate Buffered Saline: PBS) e incubados em estufa a 37[degrees]C durante uma hora, utilizando o cromogeno vermelho VECTOR[R] NovaREDD(tm).
Thus, we improved sensitivity of the test by using Labeled Streptavidin Biotin (LSAB