ELISA PROTEIN EXTRACTION, ISOLATION & SOLUBILIZATION BUFFER FOR ELISA SAMPLES (Part No. ELSP-1)

ELISA PROTEIN EXTRACTION, ISOLATION & SOLUBILIZATION BUFFER

60 ml, 1X,  pH 7.4 mild detergent protein extraction buffer that maintains antibody-antigen interactions required for ELISA sample isolation.  Extracts proteins from most cellular compartment, including membranes, cytoplasm and organelles for ELISA sample preparation.  

Highlights

• Validated formulation for ELISA sample preparation.
• Widely applicable for most cytoplasmic, transmembrane and extracellular proteins for ELISA sample preparatoin.
• Convenient pre-mixed 1X solution.
• Compatible with protein concentration assays such as Bradford.

  Contents                                                                                                                                      

1. 60 ml ELISA protein extraction buffer solution pH 7.4
   

View Protocol

The ELISA protein extraction buffer contains a mild detergent enabling desolution of cell membranes, solubilization of most membrane proteins and suspension of cytosolic proteins into the buffer.

  Protocol Overview:  ELISA Protein Extraction Buffer                                                                     

1. Prepare 500 µl ELISA protein extraction buffer for each 10-cm dish, or 200 µl for each well in a 6-well dish.  Make the solution ice cold and add protease inhibitors, including serine protease inhibitor (such as PMSF).  If needed, also add general phosphatase inhibitor.
   
2. Remove media and rinse cells three times with PBS.
3. Add 500 µl ELISA protein extraction buffer into each 10-cm dish.  Rotate the dish to cover cells with ELISA protein extraction buffer and place on a bed of ice for 5 min.
4. Scrape cells off the dish, and transfer to an appropriate tube.  Rotate and tap the tube several times to solubilize membranes.  You should observe fibrous materials which correspond to genomic DNA, an indicator of cell lysis.  If fibrous materials are not apparent, add 20 µl aliquots of ELISA protein extraction buffer, and rotate and tap until the fibrous materials are observed.  Keep the tube on ice for another 15 min. 
5. Centrifuge the tube at 4^oC for 15min and collect the supernatant, which will be used in the ELISA assay.   A protein concentration assay, such as the Bradford Assay, can be used to quantify total protein concentration.  The supernatant can also be stored at -80^oC long term