User:Wilfred J. Poppinga/Notebook/cAMP compartmentalization/2010/04/15

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 * style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]] SDS-PAGE and Immunoblot
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 * style="background-color: #F2F2F2" align="center"|  |Main project page


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Summary

 * SDS-PAGE
 * 2x 8% gel with Spectra™ Multicolor High Range Protein Ladder (Fermentas) [[Image:Spectra_Multicolor_High_Range_Protein_Ladder_-_Fermentas.jpg]]
 * 1x 15% gel with Precision Plus Protein Dual Color marker [[Image:Biorad_-_Precision_Plus_Protein_Dual_Color_marker.jpg|right|Biorad Precision Plus Protein Dual Color marker]]

Method

 * 8% (2x) and 15% (1x) gels were prepared (14April2010 as described 01March2010
 * Gels were run and transferred as described 09April2010
 * Membranes were treated with Ponceau S staining and blocked in 5% BSA TBST solution for 1 h @ RT
 * Antibodies used on membranes
 * AKAP7(Rabbit polyclonal antibody) (Abnovation)
 * 15% gel membrane (below 25 kDa)
 * GSKIP (Rabbit) (In House)
 * Lower half of 15% gel membrane (below 25 kDa)
 * AKAP-Lbc (Goat polyclonal IgG) (Santa Cruz Biotechnology)
 * 8%
 * AKAP18δ5 (Rabbit) (In House)
 * 15%
 * AKAP95 (Mouse) (Transuction Laboraties)
 * 8%
 * AKAP12 (Sheep polyclonal) (Abcam)
 * 8%
 * AKAP450 (Mouse) BD Transduction
 * 8%
 * AKAP1 (Rabbit) (Bethyl)
 * 8%
 * mAKAP (Rabbit)(Upstate)
 * 8%
 * AKAP79 (Rabbit)(Upstate)
 * Upper half of 15% gel membrane (above 25 kDa)

Discussion

 * ✅ Ow..

Attachment
   [[Media:FILE.xls| Excel TITLE]]


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