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MEDOX-BIOTM Dnase I Foot Print Analysis Of Protein-DNA Binding Teaching Kit

MEDOX-BIOTM Methylation Interfrerence Assay Teaching Kit (For Analysis of Protein DNA Interaction)
MEDOX-BIOTM Uracil Interference Assay Teaching Kit (For Analysis of Protein DNA Interaction)
MEDOX-BIOTM Purification Of DNA Binding Protein Teaching Kit (Using Biotin-streptavidin Affinity System)
MEDOX-BIOTM Determination Of Protein-DNA Sequence Specificity Teaching Kit (By PCR Assisted Binding Site Selection)
 
MEDOX-BIOTM Dnase I Foot Print Analysis Of Protein - DNA Binding Teaching Kit

Rat (rGH) is expressed exclusively in cells from the anterior lobe of the pituitary gland. Using DNAsel footprinting assays, we have examined both pituitary and nonpituitary cell nuclear extracts for proteins which bind specifically to the rGH promoter and 5'-flanking region. In agreement with previous studies, we have located binding sites between -96 and -65, and between -148 and -118 for proteins which have been termed GC1 and GC2, respectively. The GC2 footprint is found using extracts from both pituitary and nonpituitary cells, but GC1 is observed only in pituitary cells.

Ordering Information

Cat.No

Product

Units

MX-1306-01

MEDOX-BIOTM DNASE 1 Foot Print Analysis of
Protein - DNA Binding Teaching Kit

5 Exps.

MX-1306-02

MEDOX-BIOTM DNASE 1 Foot Print Analysis of
Protein - DNA Binding Teaching Kit

10 Exps.

 

 

MEDOX-BIOTM Methylation Interfrerence Assay Teaching Kit (For Analysis of Protein DNA Interaction)

The methylation interference assay is the simpler of the two, involving a chemical modification of Guanines and Adenines with Dimethylsulfate to produce N-7 methyl G or N-3 methyl residues.
These residues are subject to cleavage by piperidine. The complexity of this method is somewhat increased by the need to isolate an end labeled probe.

Ordering Information

Cat.No

Product

Units

MX-1307-01

MEDOX-BIOTM Methylation Interference Assay Teaching Kit (for Analysis of Protein DNA Interaction)

5 Exps.

MX-1307-02

MEDOX-BIOTM Methylation Interference Assay Teaching Kit (for Analysis of Protein DNA Interaction)

10 Exps.

 

 

MEDOX-BIOTM Uracil Interference Assay Teaching Kit (For Analysis of Protein DNA Interaction)

In the Uracil interference analysis DNA is synthesized in the presence of dUTP to incorporate Uracil residues in place of thymine, at a rate of 0.5-1 thymine substitutions per molecule. This can be accomplished by PCR with one
labeled primer, thus probe generation may be easier than for methylation interference. Cleavage at Uracil residues requires a two step procedure, in which Uracil glycosylase removes the Uracil base, creating apyrimidinic sites which are then cleaved by piperidine

Ordering Information

Cat.No

Product

Units

MX-1308-01

MEDOX-BIOTM Uracil Interference Assay Teaching Kit
(for Analysis of Protein DNA Interaction)

5 Exps.

MX-1308-02

MEDOX-BIOTM Uracil Interference Assay Teaching Kit
(for Analysis of Protein DNA Interaction)

10 Exps.

 

 

MEDOX-BIOTM Purification Of DNA Binding Protein Teaching Kit (Using Biotin-streptavidin Affinity System)

A method for the rapid identification and purification of se-quence specific DNA binding proteins based on magnetic separation is presented. This method was applied to confirm the binding of the human recombinant USF1 protein to its putative binding site (E-box) within the human SOX3 protomer. It has been shown that biotinylated DNA attached to streptavidin magnetic particles specifically binds the USF1 protein in the presence of competitor DNA. It has also been demonstrated that the protein could be successfully eluted from the beads, in high yield and with restored DNAbinding activity. The advantage of these procedures is that they could be applied for the identification and purification of any high-affinity sequence-specific DNA binding protein with only minor modifications.

Ordering Information

Cat.No

Product

Units

MX-1309-01

MEDOX-BIOTM Purification of DNA Binding Protein Teaching Kit (Using Biotin-streptavidin Affinity System)

5 Exps.

MX-1309-02

MEDOX-BIOTM Purification of DNA Binding Protein Teaching Kit (Using Biotin-streptavidin Affinity System)

10 Exps.

 

 

MEDOX-BIOTM Determination Of Protein-DNA Sequence Specificity Teaching Kit (By PCR Assisted Binding Site Selection)

Limited information exists for the binding specificities of many important transcription factors. To address this, we have previously developed a microwell-based assay for directly measuring the affinity of DNA-protein binding interactions. The detailed protocol for determining sequence specificities of DNA-binding proteins using this assay. The described method is rapid; after preparation of the reagents, the assay can be run in a single day, and its throughput can be increased further by automation.
The method is quantitative but requires prior knowledge of one high-affinity binding site for the protein of interest. The protocol can be adapted for determining the effect of protein modifications and protein-protein interactions on DNA-binding specificity, and for engineering proteins with new DNA-binding specificities. In addition, the method is suitable for high-throughput screening to identify proteins or small molecules that modulate protein-DNA binding interactions.

Ordering Information

Cat.No

Product

Units

MX-1310-01

MEDOX-BIOTM Determination of Protein- DNA Sequence
Specificity Teaching Kit (by PCR Assisted Binding Site Selection)

5 Exps.

MX-1310-02

MEDOX-BIOTM Determination of Protein- DNA Sequence
Specificity Teaching Kit (by PCR Assisted Binding Site Selection)

10 Exps.

 

 
   
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