2 results about "Merokeratin" patented technology

The application relates to the technical field of biology, and discloses a method for inducing a cell line of immortalized musk deer scent glands to synthesize dihydrotestosterone, wherein the cell line is obtained by transfecting a primary musk deer scent gland epithelial cell with a lentivirus carrying an SV40 Large T antigen and performing resistance screening; the cell line is in an epithelial cell form and stably expresses keratin 18, cholesterol side chain cleavage enzyme, 17 alpha-hydroxylase and 3 beta-hydroxysteroid dehydrogenase. Through lentivirus-mediated SV40 Large T antigen transfection, the primary musk deer scent gland epithelial cell breaks through the limit of replicative senescence and obtains unlimited proliferation capacity, effectively overcomes the defects that the primary cell is prone to fibrosis and rapidly loses metabolic function during in-vitro culture, provides a standardized cell model with a clear genetic background and persistent metabolic activity for biological manufacturing of dihydrotestosterone, and relieves the dependence on endangered animal live resources.

Abstract A method for detecting a biological marker in a sample, preferably a biological marker on a cancer cell, the method comprising: (i) contacting the sample with at least one DNA aptamer coupled to a first reagent and wherein the aptamer specifically binds to a first biological marker in the sample to form a first complex; (ii) contacting the first complex of (i) with a second binding agent that specifically binds to the first reagent such that a second complex is formed, wherein the second binding agent is coupled to a reporter molecule; (iii) contacting the second complex (ii) with a substrate for the reporter molecule of the at least one second binding agent; and (iv) detecting the substrate reaction by formation of a reaction product thereby detecting the biological marker in the sample. Biological markers include cytokeratin 8 (CK8), cytokeratin 18 (CK18), human epidermal growth factor receptor 2 (HER2) and epithelial cell adhesion molecule (EpCAM). Abstract 41 / 50 FIGURE 40 Frozen section preparation 4 Sample fixation + Wash (10 dips in PBS) DNA aptamer staining DAB Substrate DAB substrate HRP FAM Anti-FITC conjugated with HRP FAM DNA Aptamer conjugated with FAM HRP Frozen section slide / Touch imprint slide / Cytology slide / Direct smear slide Anti-FITC-HRP Wash (10 dips in PBS) DAB Wash (10 dips in PBS) Hematoxylin counter-staining Wash under tap water Cove rs lip DAB brown precipitate at the location of the aptamer B o min 1 min 15 sec 5 min 15 sec 5 min 15 sec 5 min 15 sec 10 sec 1 min 1 min A + Wash (10 dips in PBS) Total time for aptamer-DAB staining: —18 min 4 1 / 5 0 A B 1 0 m i n 1 m i n D A B s u b s t r a t e 1 5 s e c H R P A n t i - F I T C 5 m i n D N A a p t a m e r s t a i n i n g H R P 1 5 s e c 5 m i n 1 5 s e c D N A A p t a m e r c o n j u g a t e d w i t h F A M 5 m i n D A B W a s h ( 1 0 d i p s i n P B S ) F r o z e n s e c t i o n s l i d e / T o u c h i m p r i n t s l i d e / C y t o l o g y s l i d e / D i r e c t s m e a r s l i d e c o u n t e r - s t a i n i n g 1 m i n W a s h u n d e r t a p w a t e r 1 m i n C o v e r s l i p F I G U R E 4 020 25 28 36 33 19 D ec 2 02 5 1 9 D e c 2 0 2 5 A B 1 0 m i n F r o z e n s e c t i o n p r e p a r a t i o n 1 m i n S a m p l e f i x a t i o n D A B 2 0 2 5 2 8 3 6 3 3 s u b s t r a t e 1 5 s e c W a s h ( 1 0 d i p s i n P B S ) H R P A n t i - F I T C 5 m i n D N A a p t a m e r s t a i n i n g H R P 1 5 s e c 5 m i n A n t i - F I T C - H R P T o t a l t i m e f o r 1 5 s e c D N A A p t a m e r W a s h ( 1 0 d i p s i n P B S ) c o n j u g a t e d w i t h F A M 5 m i n D A B s t a i n i n g : ~ 1 8 m i n W a s h ( 1 0 d i p s i n P B S ) F r o z e n s e c t i o n s l i d e / T o u c h i m p r i n t s l i d e / 1 0 s e c H e m a t o x y l i n C y t o l o g y s l i d e / D i r e c t s m e a r s l i d e c o u n t e r - s t a i n i n g 1 m i n W a s h u n d e r t a p w a t e r 1 m i n C o v e r s l i p F I G U R E 4 0