Indlela yokwenza le ncwadana ye-Immunohistochemical Staining Buffer
I-phosphate i-saline ehluthayo (PBS) isisombululo samathambo esetyenziswa ngokuqhelekileyo kwi-immunohistochemical (IHC) kunye nokuhlala isetyenziswe kuphando lwezinto eziphilayo. I-PBS isisombululo setyuwa samanzi esine-sodium hydrogen phosphate, i-sodium chloride kwaye, kwezinye iimeko, i-potassium chloride kunye ne-potassium dihydrogen phosphate.
I-Immunohistochemical Staining
I-Immunohistochemistry ibhekisela kwinkqubo yokufumanisa ama-antigen njengama-proteins kwiiseli zesigaba seesisu ngokusebenzisa umgaqo we-antibodies ezibophelela ngokuthe ngqo kuma-antigen kwiimpawu eziphilayo.
I-stain immunofluorescent yayiyindlela yokuqala yokugcoba i-immunohistochemical.
Ama-antigens abonakalayo xa adibaniswa nama-antibodies usebenzisa idayi ye-fluorescence ngenxa yempendulo ye-antigen-antibody yokuzibophezela. Le nqubo iyenzeka xa isebenze ngokukhanya okuthakazelisayo kwinqanaba elithile elingaphantsi kwe-microscope ye-fluorescent.
I-osmolarity kunye ne-ion concentrations yezisombululo ezifanisana nomzimba womntu-ziyi-isotonic.
I Recipe yePBS
Unokulungiselela iPBS ngeendlela ezininzi. Kukho amafomula amaninzi. Ezinye zazo aziqukethe i-potassium, ngelixa ezinye ziqukethe i-calcium okanye i- magnesium .
Le recipe ilula. I-10X ye-stock PBS isisombululo (0.1M). Nangona kunjalo, unokwenza isisombululo se-1X, okanye uqale nale recipe ye-10X uze uyilungise kwi-1X.
Inkqubo yonke ithatha malunga nemizuzu eyi-10.
Inketho yokongeza i-Tween ikwabonelelwa kwakhona.
Oko Uya Kufuna Ukwenza I-PBS Buffer
- I-sodium phosphate monobasic (i-anhydr)
- I-sodium phosphate dibasic (i-anhydr)
- I-chloride ye sodium
- Isilinganiselo kwaye ulinganise izikebhe
- Umqhubi wamagnetic kwaye ugxobhoze ibha
- Inkqubo ye-pH efanelekileyo kunye neendlela ezifanelekileyo zokulungisa i-pH
- I-1L ibhola lokuvutha
- Uphakathi kwe-20 (ukhetho)
Indlela yokwenza i-Phosphate Buffered Saline (PBS)
I-10.9g ye-sodium phosphate dibasic (Na2HPO4), 3.2g ye-sodium iphosphate monobasic (NaH2PO4) kunye ne-90g ye-chloride ye-sodium (NaCl).
Dissolve nje ngaphantsi kwe-1L amanzi adibeneyo.
Lungisa i-pH ukuya kwi-7.4 uze wenze isisombululo ukuya kumqulu wokugqibela we-1L.
Nciphisa i-10X ngaphambi kokusetyenziswa kwaye ulungise i-pH xa kuyimfuneko.
Unokwenza isisombululo sePBS esine-0.5 yepesenti phakathi kwe-20 ngokufaka i-5mL Tween 20 kwisisombululo se-1L.
Iingcebiso zokwenza i-PBS Buffer
Gcina i-tampu kwindawo yokushisa egumbini emva kokuba wenze isisombululo sePBS.
Ama-reagents angenayo i-anhydr can substitute but you need to recalculate the mass of each one to accommodate the molecules.
Ukusetyenziswa kwe-Phosphate Buffered Saline (PBS)
I-phosphate i-saline ephuhliweyo ine-usetyenziso emininzi kuba i-isotonic kwaye ayinayo isetyhi kwiiseli ezininzi. Ingasetyenziselwa ukunciphisa izinto kwaye isoloko isetyenziselwa ukuhlamba izitya zeeseli. I-PBS ingasetyenziselwa njenge-dilutent kwiindlela ezahlukahlukeneyo zokomisa i-biomolecules kuba iamolekyu zamanzi ngaphakathi kwayo ziya kulungelelaniswa malunga neprotheni ye-substance, umzekelo. Kuya "kumiswa" kwaye kungenakunqandwa kummandla oqinileyo.
I-pH ihlala ihleli kwaye ihambelana nokuthintela ukutshatyalaliswa kweeseli.
Ifilimu emancinci yamanzi ebophezelayo kwizinto ezinqanda i-denaturation okanye ezinye iinguqu eziguqukayo. Iibhobho zokunqanda i-Carbonate zingasetyenziselwa injongo enye kodwa ngezinga elincinci.
I-PBS ingasetyenziselwa ukuthatha i-spectrum xa ilinganisa i- protein ye- adsorption kwi-ellipsometry.