To deplete Wif1 from OPC conditioned medium, APCfl/fl control or Olig2cre:APC fl/fl OPC conditioned medium were incubated with Wif1 antibody (1:100, rabbit, ab155101, Abcam) at 4℃ overnight followed by pull down with agarose beads (sc-2003, Santa Cruz). BSA was used as the non-antibody control. The purified Wif1-depleted conditioned media were then used for endothelial cell treatment. To clarify the concentrations of Wif1 protein in the conditioned media before and after Wif1 depletion, four groups of conditioned media (WT-CM+BSA, Olig2cre:APC fl/fl-CM+BSA, WT-CM+Ab and Olig2cre:APC fl/fl-CM+Ab) were examined using the Wif1 specific ELISA kit (mouse WIF1 PicoKine Elisa Kit, EK1523, Boster) according to the manufacturer’s instructions. The OD values were determined by measuring the absorbance at 450nm using the microplate reader (Bio-RAD, Model 680). Independent experiments were performed in triplicate.
Mouse wif1 picokine elisa kit
Manufactured by Boster Bio
The Mouse WIF1 PicoKine ELISA Kit is a quantitative sandwich enzyme-linked immunosorbent assay (ELISA) designed for the accurate measurement of mouse WIF1 protein levels in cell culture supernatants, cell lysates, and other biological fluids.
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2 protocols using mouse wif1 picokine elisa kit
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Wif1 Depletion from OPC Conditioned Media
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Wif1 Depletion from OPC Conditioned Media
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To deplete Wif1 from OPC conditioned medium, APCfl/fl control or Olig2cre:APC fl/fl OPC conditioned medium were incubated with Wif1 antibody (1:100, rabbit, ab155101, Abcam) at 4℃ overnight followed by pull down with agarose beads (sc-2003, Santa Cruz). BSA was used as the non-antibody control. The purified Wif1-depleted conditioned media were then used for endothelial cell treatment. To clarify the concentrations of Wif1 protein in the conditioned media before and after Wif1 depletion, four groups of conditioned media (WT-CM+BSA, Olig2cre:APC fl/fl-CM+BSA, WT-CM+Ab and Olig2cre:APC fl/fl-CM+Ab) were examined using the Wif1 specific ELISA kit (mouse WIF1 PicoKine Elisa Kit, EK1523, Boster) according to the manufacturer’s instructions. The OD values were determined by measuring the absorbance at 450nm using the microplate reader (Bio-RAD, Model 680). Independent experiments were performed in triplicate.
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