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This study focuses on optimizing immunofluorescence and Western blot protocols for observing growth in untreated A498 spheroid cells. Concentration adjustments for FLT-1, PDGFR-α, and PDGFR-β markers were analyzed. Additionally, Western blot optimizations were performed for p70S6K and 4E-BP1 proteins.
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IF 3 EJ28 – Formalinfixierung 1° AK: 1:50 2° AK: 1:30 neg. control FLT1 PDGFR-α
IF 3 A498 – Formalinfixierung 1° AK: 1:50 2° AK: 1:30 neg.control FLT1 PDGFR-α
FLT-1 neg. contr. IF4 FLT-1 Konzentrationsoptimierung FLT-1 1:25 FLT-1 1:50 FLT-1 1:33
PDGFR-α neg. contr. IF5 PDGFR-αKonzentrationsoptimierung PDGFR-α 1:25 PDGFR-α 1zu50 PDGFR-α 1zu33
PDGFR-β neg. contr. IF5 PDGFR-βKonzentrationsoptimierung PDGFR-β 1:25 PDGFR-β 1:50 PDGFR-β 1:33
Optimierung Western Blot p70S6K 70-85 kDa, rabbit P-p70S6K 70-85 kDa, rabbit; P-Thr389 4E-BP1 15-20 kDa, rabbit P-4E-BP1 15-20 kDa, rabbit; P-Thr70