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Catalogue Number
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160878 |
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Applications
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N/A
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Antigen/Gene or Protein Targets
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p27kip1 (encoded by the CDKN1B gene)
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Reactivity
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N/A
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Relevance
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Breast cancer
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Host
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Mouse
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Positive Control
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MCF7 or Cal51 cells
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Recommended Growing Conditions
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DMEM or RPMI1640 + 10% foetal calf serum
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Notes
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Purified GST-p27kip1 fusion protein in PBS emulsified with an equal volume of Freund's complete adjuvant was injected intraperitoneally into five mice (50g/mouse). The same amount of protein in PBS emulsified with an equal volume of Freund's incomplete adjuvant was subsequently used to boost the mice at intervals of 21 days. After three injections, the titres of the sera were monitored by immunoblotting, and the mouse with the highest titre was injected intraperitoneally with 50g of GST-p27kip1 protein in PBS only. On Day 3 after the final boost, the mouse was killed and the spleen was removed. The spleen cells were fused with mouse myeloma cells Sp2/o-Ag14 using 50% polyethylene glycol 1500. The cells were grown in Dulbecco's minimal essential medium containing 20% foetal calf serum supplemented with interleukin-6 at 4ng/ml. The hybridoma cells were selected using hypoxanthine and azaserine.
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Research Area
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Cancer
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