Western Blot Analysis of Prostate Samples

Protein extraction from frozen prostate samples for Western blot analysis has previously been described [49 (link)]. Western blots were prepared using 15–20 µg of protein lysate, blocked with 5% BSA in 1 × TBS / 0.1% Tween-20 for 1 h and incubated at 4 °C overnight with primary antibodies against AR (1:1000, ab133273, Abcam), p16^INK4A (1:2000, ab211542, Abcam), CDK4 (1:200, sc-260, Santa Cruz), p53 (1:1000, CST#2524, Cell Signaling), β-Actin (1:1000, CST#4967, Cell Signaling) and β-Tubulin (1:2000, CST#2146, Cell Signaling). Western blots were quantified using ImageJ2.

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Limberger T., Schlederer M., Trachtová K., Garces de los Fayos Alonso I., Yang J., Högler S., Sternberg C., Bystry V., Oppelt J., Tichý B., Schmeidl M., Kodajova P., Jäger A., Neubauer H.A., Oberhuber M., Schmalzbauer B.S., Pospisilova S., Dolznig H., Wadsak W., Culig Z., Turner S.D., Egger G., Lagger S, & Kenner L. (2022). KMT2C methyltransferase domain regulated INK4A expression suppresses prostate cancer metastasis. Molecular Cancer, 21, 89.

Publication 2022

ab133273 ab211542 sc-260 CST#2524 β-Actin CST#4967 β-Tubulin CST#2146

Actin Antibodies Frozen P16ink4a Prostate Protein Tubulin Tween 20 Western blots

Corresponding Organization :

Other organizations : Medical University of Vienna, Central European Institute of Technology – Masaryk University, Central European Institute of Technology, University of Veterinary Medicine Vienna, Center For Biomarker Research In Medicine, Innsbruck Medical University, Ludwig Boltzmann Institute Applied Diagnostics

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Variable analysis

independent variables

Primary antibodies against AR, p16^INK4A, CDK4, p53, β-Actin, and β-Tubulin

dependent variables

Protein expression levels of AR, p16^INK4A, CDK4, p53, β-Actin, and β-Tubulin

control variables

Protein lysate amount (15-20 μg)
Blocking buffer (5% BSA in 1 × TBS / 0.1% Tween-20)
Incubation time (1 h)
Incubation temperature (4 °C overnight)

controls

Positive controls not explicitly mentioned
Negative controls not explicitly mentioned

Annotations

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