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Immunofluorescence and Western Blot Optimization Study on Growth Observation in Untreated Spheroid A498 Cells

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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Immunofluorescence and Western Blot Optimization Study on Growth Observation in Untreated Spheroid A498 Cells

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  1. Größenbeobachtung A498 Sphäroide- unbehandelt

  2. 1. RAD001 Behandlung

  3. IF 3 EJ28 – Formalinfixierung 1° AK: 1:50 2° AK: 1:30 neg. control FLT1 PDGFR-α

  4. IF 3 A498 – Formalinfixierung 1° AK: 1:50 2° AK: 1:30 neg.control FLT1 PDGFR-α

  5. FLT-1 neg. contr. IF4 FLT-1 Konzentrationsoptimierung FLT-1 1:25 FLT-1 1:50 FLT-1 1:33

  6. PDGFR-α neg. contr. IF5 PDGFR-αKonzentrationsoptimierung PDGFR-α 1:25 PDGFR-α 1zu50 PDGFR-α 1zu33

  7. PDGFR-β neg. contr. IF5 PDGFR-βKonzentrationsoptimierung PDGFR-β 1:25 PDGFR-β 1:50 PDGFR-β 1:33

  8. PDGFR-β 1:25 FITC und DAPI kombiniert

  9. 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

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