Navigation Links
Total Cellular Protein Determination Using the DC Protein Assay


Introduction
The DC Protein Assay reagents contain both detergent and base which are required for cell lysis of many cell samples prior to determination of total cellular protein. The following protocol can be used for tissue cells; dry cells; protein precipitates from heat, acid, or acetone treatment; or any other samples that require both sodium hydroxide and detergent for solubilization.


Protocol
1. Wash cells or protein precipitate. Using an isotonic solution such as TBS or PBS, suspend the sample in excess buffer and mix by inversion or vortex mixer. Centrifuge sample or precipitate until a tight pellet has formed. Discard the supernatant.

2. Prepare working reagent A' by mixing 20 l Reagent S to each milliliter of Reagent A that will be needed for the assay. Mix by gentle inversion to prevent foaming.

3. Add 1 ml working reagent A' to approximately 1 x 106 cells. Vortex intermittently for 510 minutes, and heat to 60 C (if necessary) to solubilize the pellet.

4. Cool mixture to room temperature. In order to accurately determine protein concentration, several dilutions of the digestion with reagent A' should be made (1:2, 1:4, 1:8, etc.). Record the dilution factors for correction in the final protein concentrations.

5. Create a standard curve. Transfer 100 l of the standard into a clean test tube. Three to five data points should be generated. Either standard supplied with the kit (BSA or IgG) can be used, since both react similarly to the DC Protein Assay reagents.

6. Transfer 500 l sample of serially diluted samples from step 5 into clean test tubes.* To correct for volume difference between samples and standard curve, add 100 l water to each tube. Add 4.0 ml Reagent B and vortex.

7. Add 500 l working reagent A' to samples in the standard curve series and vortex.

8. Add 4.0 ml Reagent B, and vortex.

9. Incubate standards and samples at room temperature for 15 minutes. Measure absorbance at 650750 nm. Compare results to a standard curve.


For technical assistance, call your local Bio-Rad representative, or in the U.S., call 1-800-4BIORAD.


back to top
'"/>

Source:


Page: All 1 2

Related biology technology :

1. New Kit Generates Exceptionally Pure Total RNA for RT-PCR
2. High-Throughput Isolation of Total RNA
3. Isolate and Analyze Total RNA from Cells Harvested by Laser Capture Microdissection
4. Fast Isolation of Total RNA from Small Numbers of Cells
5. Total RNA Isolation Protocol 1,2,3
6. Isolate It All: siRNA miRNA Total RNA Native Protein
7. Isolate Total RNA and Protein From the Same Sample
8. Isolate High Quality Total RNA from LCM Samples Suitable for Microarray and qRT-PCR Analysis
9. Recover High Yields of Total Nucleic Acid from Formalin-fixed, Paraffin-embedded (FFPE) Tissue
10. Total RNA from Whole Blood for Expression Profiling
11. RNAPrep - Total RNA Isolation Purification
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:5/27/2016)... ... , ... Weeks after hosting a carpal tunnel syndrome workshop with Dr. Oz ... founder of the Fitzmaurice Hand Institute, has announced the addition of MRI diagnostic imaging ... and only 1 of about 3 currently available in the United States. Developed specifically ...
(Date:5/27/2016)... 27, 2016 At present, the Biotech ... this space know that volatility is what makes this industry ... ActiveWallSt.com: Synta Pharmaceuticals Corp. (NASDAQ: SNTA ), CTI ... LPTN ), and Heat Biologics Inc. (NASDAQ: ... the technical alerts for these stocks at: ...
(Date:5/26/2016)... -- Despite the volatility that continues to envelop ... pre-market research on ActiveWallSt.com directs the investor community,s focus on ... RDUS ), Cerus Corp. (NASDAQ: CERS ), Arrowhead ... Therapeutics Inc. (NASDAQ: FPRX ). Register with us ... http://www.activewallst.com/ On Wednesday, shares in ...
(Date:5/25/2016)... ... ... Lajollacooks4u has become a rising hotspot for specialized team building events in ... Fortune 500 companies, such as Illumina, Hewlett-Packard, Qualcomm and Elsevier, have traveled from ... Each event kicks off with an olive oil and salt-tasting competition. From there, ...
Breaking Biology Technology:
(Date:4/14/2016)... , April 14, 2016 ... and Malware Detection, today announced the appointment of ... the new role. Goldwerger,s leadership appointment comes ... the heels of the deployment of its platform at ... behavioral biometric technology, which discerns unique cognitive and physiological ...
(Date:3/23/2016)... 23, 2016 ... Gesichts- und Stimmerkennung mit Passwörtern     ... MESG ), ein führender Anbieter digitaler ... mit SpeechPro zusammenarbeitet, um erstmals dessen Biometrietechnologie ... die Möglichkeit angeboten, im Rahmen mobiler Apps ...
(Date:3/17/2016)... , March 17, 2016 ABI Research, ... forecasts the global biometrics market will reach more ... 118% increase from 2015. Consumer electronics, particularly smartphones, ... fingerprint sensors anticipated to reach two billion shipments ... Dimitrios Pavlakis , Research Analyst at ABI ...
Breaking Biology News(10 mins):