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
'"/>

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:
TAG: Total Cellular Protein Determination Using the Protein Assay

(Date:1/14/2014)... DC (PRWEB) January 14, 2014 Carahsoft ... for Thursday, January 23, 2014 at 2pm EST (11am ... Knowledge.” The topic focuses on how technology can turn ... critical decisions for government agencies. The online webinar will ...
(Date:1/14/2014)... 14, 2014  3D Communications, a leading provider of strategic communications services ... media events in the United States and ... Virginia Cox , JD, is returning to the firm,s ... re-joins 3D after more than two years of service as Associate ...
(Date:1/14/2014)... NY (PRWEB) January 14, 2014 EquitiesIQ, ... Alliqua, Inc. (OTCQB: ALQA). Alliqua is an emerging biomedical ... serve the wound care market. , Free report download: ... restructured with a seasoned management team and Board, which ...
(Date:1/14/2014)... SAN DIEGO and BETHESDA, Md. ... they are joining together with two institutes from the ... more reliable tools for bringing safer, more effective treatments ... with the National Center for Advancing Translational Sciences (NCATS) ...
Breaking Biology Technology:Webcast - Natural Language Processing: Converting Raw Data into Actionable Knowledge – Hosted by Carahsoft and CDS Federal Services 2Former FDA Associate Commissioner Returns To 3D Communications 2EquitiesIQ Initiates Coverage of Alliqua, Inc. 2EquitiesIQ Initiates Coverage of Alliqua, Inc. 3Organovo Announces Collaboration with National Institutes of Health 2Organovo Announces Collaboration with National Institutes of Health 3
... Oct. 24, 2011 Marshall Edwards, Inc. (Nasdaq: ... of novel therapeutics targeting cancer metabolism, announced today that Charles ... Company, has been appointed to its board of directors. The ... to six and the number of independent directors to five. ...
... (NYSE symbol: OME), a nutritional ingredient company and the ... fish meal products, today announced that it has retained ... with its omega-3 fish oil and omega-3 concentrate research ... analytical services and biotechnology consulting company that specializes in ...
... Oct. 24, 2011 Heart repair.  Bone repair. ...  Cerebral Palsy.  Diabetes.  Alzheimer,s.  Cystic Fibrosis.  Leukemia, Sickle ... metabolic disorders. "These diseases or group ... cured with umbilical cord blood stem cells or ...
Cached Biology Technology:Marshall Edwards Appoints Healthcare Investment Banking Veteran Charles Baltic to Board of Directors 2Marshall Edwards Appoints Healthcare Investment Banking Veteran Charles Baltic to Board of Directors 3Omega Protein Announces Retention of Lipid Technologies, LLC 2Cord Blood America Provides Insight on Worldwide Research Using Umbilical Cord Blood Stem Cells to Treat Numerous Diseases 2Cord Blood America Provides Insight on Worldwide Research Using Umbilical Cord Blood Stem Cells to Treat Numerous Diseases 3
(Date:4/22/2014)... club of a mantis shrimp, a team of researchers ... University of Southern California and Purdue University, have developed ... impact resistant and tougher than the standard used in ... this tiny crustacean, the more we realize its structure ... said David Kisailus, a Kavli Fellow of the National ...
(Date:4/22/2014)... Biology (FASEB) has released updated factsheets for ... National Institutes of Health (NIH) benefits each of the ... to make these factsheets available to help citizens and ... said FASEB President, Margaret K. Offermann, MD, PhD. NIH ... funding, investing $29.2 billion in FY 2013 in medical ...
(Date:4/22/2014)... begins a trip to Asia to coordinate with ... Pacific, officials at the Office of Naval Research ... Partnership (APTEP) as an example of strong and ... allies in the region. , The APTEP program, ... region, supports the development of alternative energy technologies. ...
Breaking Biology News(10 mins):Mantis shrimp stronger than airplanes 2Mantis shrimp stronger than airplanes 3Full power: Alternative energy partnerships flourish in Asia 2Full power: Alternative energy partnerships flourish in Asia 3
... CREEK, CAMessages from nearly a half-billion years ago, conveyed ... moss, provide clues about the earliest colonization of dry ... Joint Genome Institute (DOE JGI) was among the leaders ... to complete the first genome sequencing project of a ...
... published a paper today that is the most comprehensive ... are having on the worlds coral reefs. The ... and Ocean Acidification, co-authored by seventeen marine scientists from ... not survive the drastic increases in global temperatures and ...
... The Stowers Institutes Rong Li Lab, in collaboration ... quantitative in vivo measurement of the dynamic protein-protein ... signaling pathway, which is critical to growth and ... paper, Mapping Dynamic Protein Interactions in the MAP ...
Cached Biology News:DOE JGI Community Sequencing Program delivers first moss genome 2DOE JGI Community Sequencing Program delivers first moss genome 3Immediate action needed to save corals from climate change 2Immediate action needed to save corals from climate change 3Rong Li Lab reports protein interactions of MAP kinase signaling pathway 2
Physical Form: powder Color: white Removes FLAG peptide from N-terminal and Met-N-terminal fusion proteins. Supplied with optimized enterokinase buffer....
This antibody detects ubiquitinated proteins on SDS-PAGE immunoblots. It does not detect free ubiquitin....
... The Jouan B4i ventilated and BR4i refrigerated multifunction ... low speed operation at a low sound level ... certified for biocontainment. Their AUTO-LOCK rotor exchange system ... ready for use within seconds. Instant ...
... BR4i refrigerated multifunction centrifuges are equally capable of ... low sound level with a large range of ... rotor exchange system without tools ensures that the ... Instant rotor exchange (5 sec.) ...
Biology Products: