Navigation Links
Purification and Characterization of beta-Lactoglobulin Genetic Variants A and B Using Preparative Electrophoresis and Isoelectric Focusing

ropanol/7% acetic acid, and were destained with 50% isopropanol/10% acetic acid.

ANALYTICAL IEF
The isoelectric points of the two βLG fractions separated on the Model 491 Prep Cell, were analyzed by modifying the method obtained from Bio-Rad (IEF Ready Gel Application Guide). IEF was performed on Bio-Rad IEF Ready Gels (pH 58) with 20 mM NaOH as a cathode buffer and 7 mM phosphoric acid as an anode buffer on Bio-Rads Mini Prep Cell apparatus. The two βLG fractions and the standards were dissolved in distilled water containing 10% glycerol (1 mg/ml). The run was performed at 5 W constant power and 500 Vmax. for 2.5 h. The gels were stained and destained as described above.

IMMUNOBLOTTING
The two βLG fractions, from preparative electrophoresis, were also characterized by polyclonal antiserum to βLG (Nordic Immunologic, Tilburg, Netherlands). Standards were as described above. Sample proteins, separated by PAGE or IEF, were electrophoretically blotted into nitrocellulose (0,45 m nitrocellulose membrane, Hybond-C, Amersham) by the method of Towbin.32 The electrophoretic blotting was performed with 25 mM Tris, 192 mM glycine, pH 8.3, at 30 V overnight. Residual binding sites were blocked with 1.0% bacto-gelatin for 2 h at RT, and the nitrocellulose sheet was then washed three times with PBS Tween 20 (0.05%), 10 minutes each. To detect the βLG specific proteins, the nitrocellulose sheet was incubated with the polyclonal antiserum to βLG (5 l/50 ml PBS/0.05% Tween 20) for 2 h at RT, washed three times as above, and then incubated with horseradish peroxidase-coated second antiserum (10 l/50 ml PBS Tween 20 (0.05%) for
'"/>

Source:


Page: All 1 2 3 4 5 6 7 8 9 10 11 12

Related biology technology :

1. High-Level Protein Expression, One-Column Purification, and FLAG Epitope Tagging in E. coli
2. Simple Purification of DNA from Plasmid Minipreps, PCR Amplifications and Agarose Gels
3. Yeast Protein Production System Features High Yields and One-Step Purification
4. Expression and Purification of Recombinant Proteins That Have Native Amino Acid Sequence
5. Easy Cosmid DNA Purification with StrataPrep Kits
6. MinElute PCR Purification Kit
7. QIAquick PCR Purification Kit
8. Microarray Target Purification Kit and the Microarray Target Synthesis Kit (T7)
9. Double-Affinity Purification of GST-His Tagged Proteins Using Vivapure 8-to-96 Well Cobalt-Chelate Kit
10. Ultra-Fast Purification of Small Nucleic Acids
11. RNAClean - RNA & cDNA in vitro Reaction Purification
Post Your Comments:
(Date:8/26/2014)... The 20 ambassadors representing the ALS Therapy ... the following statement on Facebook today regarding the global ... funding specifically for ALS research: , “Who would have ... would raise all this awareness and funding toward ALS ... behalf of the entire ALS community, would like to ...
(Date:8/26/2014)... argument has just been added to the growing ... as the next big thing in the high-tech ... materials. An international collaboration of researchers led by ... (DOE),s Lawrence Berkeley National Laboratory (Berkeley Lab) has ... transfer in photo-excited MX2 materials. The recorded charge ...
(Date:8/26/2014)... , Aug. 26, 2014 An analysis by ... of all Dengue virus strains in Pubmed, has revealed ... to 1944. This conservation and sharing of specific Replikin ... Dengue Blocker-Vaccine™ candidate, as it did for the H5N1 ... of which have been found to be effective (1,2). ...
(Date:8/26/2014)... Mich. , Aug. 26, 2014   ... development of breakthrough oral medicines to treat autoimmune ... immune modulation, today announced the appointment of ... company,s scientific advisory board.  Dr. Singh will lead ... to advance Lycera,s multiple ongoing research programs and ...
Breaking Biology Technology:ALS TDI’s Young Faces of ALS Ambassadors Thank You for Accepting the Ice Bucket Challenge to End ALS 2ALS TDI’s Young Faces of ALS Ambassadors Thank You for Accepting the Ice Bucket Challenge to End ALS 3ALS TDI’s Young Faces of ALS Ambassadors Thank You for Accepting the Ice Bucket Challenge to End ALS 4Competition for graphene 2Lycera Appoints Harinder Singh, Ph.D., as Chair of Scientific Advisory Board 2Lycera Appoints Harinder Singh, Ph.D., as Chair of Scientific Advisory Board 3
... - Educators and software developers will have the ... learning process at the first annual Games+Learning+Society conference ... conference is being held by Games And Professional Practice ... Learning (ADL) Co-Lab and a group of instructors at ...
... Marquette University and UGS, a provider ... the largest in-kind contribution ever presented to Marquette University, in ... a market value it says is more than $30.5 million. ... Jim Factor's Geometric Modeling Lab in the Department of Mathematics, ...
... Assembly on Thursday passed AB 499, a proposed ban on ... would outlaw the cloning of embryos both for reproductive purposes ... the making of a cloned embryo for the purpose of ... were to become law would go up to ten years ...
Cached Biology Technology:Education conference to draw together games researchers 2
(Date:8/25/2014)... fires began as a lightning strike on August 12, 2014. ... 20% contained. There are close to 2,000 personnel ... many small fires and a number of large ones. ... account for about 100 acres. An inversion layer settled ... on August 24, keeping fire activity moderate. As the inversion ...
(Date:8/25/2014)... Many Americans need extraordinary willpower to avoid becoming obese ... too much. For members of minority groups, maintaining a ... new research led by Luis Rivera, an experimental social ... common for minorities in the United States to endure ... inferior, and that these attitudes can prevent people from ...
(Date:8/25/2014)... 2014 Nearly 1 in 6 adults worldwide may ... disorder can cause serious complications. Auriculotherapy, a form ... the outer ear, may help in managing constipation. ... that evaluated the effectiveness of auriculotherapy in treating ... a Review article in The Journal of ...
Breaking Biology News(10 mins):Happy Camper and July Fire Complexes in California 2Racial and ethnic stereotypes may contribute to obesity among minorities 2
... its damaging effect on the environment and its illegal ... to adverse effects on human health, such as inflammation ... damage and possibly cancer. In a ... journal Environmental Research Letters, researchers took air ...
... pinpoint a small RNA that spurs cells to manufacture a ... Alzheimer,s disease (AD) or other types of neurodegeneration. The study ... of Cell Biology ( www.jcb.org ). Like ... in more than one version. Although scientists have identified numerous ...
... Nature a new molecular pathway used to suppress blood ... potential therapeutic value for fighting diseases of the retina and ... blood cells involved in the immune system, use this molecular ... in the same study researchers were able to reverse this ...
Cached Biology News:'E-waste pollution' a threat to human health, new research suggests 2Novel pathway regulating angiogenesis may fight retinal disease, cancers 2
... rapid analytical to medium scale affinity purification ... vectors. Sufficient reagents are provided for purification ... under native or denaturing conditions. Crude samples ... retains STag fusion proteins. After unbound proteins ...
High affinity antibody reacting with S-100 ab heterodimer. Could be used for Western blotting application and immunoassay....
... Ingredients (g/L):,Tryptone, 10 ,Yeast extract, 5 ... 0.3 ,Ferric ammonium citrate, 0.5 ,IPTG, ... water soluble, autoclavable chromogenic substrate for ... by -galactosidase generates a black insoluble ...
S-100 alpha/beta chain (8B10)...
Biology Products: