Investigation of photoinduced reactivity of Glue-FITC: 1- Absorption spectra: Glue-FITC 3µM + BSA 10µM in Tris-HCl buffer (Tris – 20mM, pH- 7.0)

Slides:

Advertisements

Similar presentations

Electrophoresis Theory

Advertisements

Group Meeting November 26 th, 2012 Derek Hernandez.

Quality Control of Product

Illumination and Filters Foundations of Microscopy Series Amanda Combs Advanced Instrumentation and Physics.

Electrophoresis PAGE Dr Gihan Gawish.

PROBING PROTEIN STRUCTURE AND INTERACTIONS USING FUNCTIONALIZED NAPHTHALIMIDES L. Kelly, B. Abraham, M. Mullan Department of Chemistry and Biochemistry,

A carboxylated Zn-phthalocyanine inhibits the fibril formation of Alzheimer’s amyloid β peptide Atsushi Nagai Dept. Laboratory Medicine Shimane University.

Katarzyna Polska, Stanisław Radzki Department of Inorganic Chemistry, Maria Curie-Skłodowska University Pl. M. C. Skłodowskiej 2, Lublin, Poland.

AMINO ACID ANALYSIS. Amino Acid Analysis  Amino acid analysis is the determination of what types of amino acids and how many of each compose a protein.

Separation of main plasma protein by using SDS-PAGE

Report Draw a scheme of the GA20OX cloning procedure.

Polyacrylamide Gel Electrophoresis of Proteins and the Enzyme-Linked Immunosorbent Assay 1.

Lecturer: David. * Reverse transcription PCR * Used to detect RNA levels * RNA is converted to cDNA by reverse transcriptase * Then it is amplified.

Polyacrylamide Gel Electrophoresis. Electrophoresis Horizontal Agarose Gels Agarose forms a gel or molecular sieve that supports the movement of small.

Lab Session 9 IUG, 2012 TMZ.

ISOLATION, SEPARATION AND DETECTION OF PROTEINS part I Michael Jelínek, Jan Šrámek Lenka Rossmeislová.

Proteomics For any given species, the space of possible biomolecules and their organization into pathways and processes is large but finite.

Figure e-1 A. Detection of anti-TIF1-γ antibody.

Quality Control of Product

ELECTROPHORETIC METHODS

Sodium Dodecyl Sulfate -Polyacryl Amide Gel Electrophoresis [SDS-PAGE]

Purification Of Proteins.

Amino Acids, Peptides, and Proteins

Lab Activity 11,12 IUG, TMZ.

Unravelling DELLA’s protein-protein interactions via co-Immunoprecipitation in wheat protoplasts Hi everyone, i’m yoni Bultinck Intern at Bayer and helping.

Volume 78, Issue 2, Pages (February 2000)

Southern Blotting.

Rapid Detection of the Epidermal Growth Factor Receptor Mutation in Non-Small-Cell Lung Cancer for Analysis of Acquired Resistance Using Molecular Beacons

Volume 12, Issue 1, Pages (January 2005)

Binding of Calcium Ions to Bacteriorhodopsin

Separation techniques

SDS-PAGE and immunoblot analysis of F4:5 progeny of a 10/81b × PI cross. SDS-PAGE and immunoblot analysis of F4:5 progeny of a 10/81b × PI

Overlay of two separate 2D gels of soybean seed proteins using Delta2D software. Overlay of two separate 2D gels of soybean seed proteins using Delta2D.

Activation-Dependent Modulation of Hyaluronate-Receptor Expression and of Hyaluronate-Avidity by Human Monocytes Johannes M. Weiss, Andreas C. Renkl,

Characterization of proteinuria and tubular protein uptake in a new model of oral L-lysine administration in rats K. Thelle, E.I. Christensen, H. Vorum,

Sodium Dodecyl Sulfate -Polyacryl Amide Gel Electrophoresis [SDS-PAGE]

TEV protease elution from cellulose is specific and provides highly purified target protein. TEV protease elution from cellulose is specific and provides.

Volume 96, Issue 8, Pages (April 2009)

TAT-mediated Delivery of LAD Restores Pyruvate Dehydrogenase Complex Activity in the Mitochondria of Patients with LAD Deficiency Matan Rapoport, Ann.

Binding of human secretory leukocyte protease inhibitor in uterine cervical mucus to immunoglobulins: pathophysiology in immunologic infertility and local.

Sodium Dodecyl Sulfate -Polyacryl Amide Gel Electrophoresis [SDS-PAGE]

Factor Va Increases the Affinity of Factor Xa for Prothrombin

Sodium Dodecyl Sulfate -Polyacryl Amide Gel Electrophoresis [SDS-PAGE]

SDS-PAGE analysis of mature seed from lines Patriot and PI

Yuan Lin, David S.W. Protter, Michael K. Rosen, Roy Parker

TIME RESOLVED SPECTROSCOPY [T.R.S.]:

Experiment Separation of proteins by using SDS-PAGE

Inhibitory Effect of β-Thujaplicin on Ultraviolet B-Induced Apoptosis in Mouse Keratinocytes Takako Baba, Hajime Nakano, Katsuto Tamai, Daisuke Sawamura,

Diagnostic tests for antibody or antigen

NikR Repressor Chemistry & Biology

Volume 12, Issue 1, Pages (January 2005)

Molecular Therapy - Nucleic Acids

Volume 11, Issue 1, Pages (January 2004)

John H. Wiessner, Linda Y. Hung, Neil S. Mandel Kidney International

Akemi Ishida-Yamamoto, Hidetoshi Takahashi, Hajime Iizuka

Apolipoprotein E is Present in Primary Localized Cutaneous Amyloidosis

IgG Autoantibodies from Bullous Pemphigoid (BP) Patients Bind Antigenic Sites on Both the Extracellular and the Intracellular Domains of the BP Antigen.

Role and identification of protein kinase A anchoring proteins in vasopressin-mediated aquaporin-2 translocation Enno Klussmann, Walter Rosenthal Kidney.

Sodium Dodecyl Sulfate -Polyacryl Amide Gel Electrophoresis [SDS-PAGE]

Fig. 2. Sera IgE reactivity patterns and the frequency of reactivity to raw and cooked shrimp extracts by Western blot. (A, C) All sera were diluted at.

Lab# 5 Western Blot BCH 462[practical].

Dept. of Chemistry, University of Toronto, Canada

Fibrinogen fragments and platelet dysfunction in uremia

Volume 18, Issue 23, Pages (December 2008)

Polyacrylamide Gel Electrophoresis (PAGE)

Expression and immunolocalization of the calpain-calpastatin system in the human oocyte Irit Ben-Aharon, M.D., Ph.D., Dalit Ben-Yosef, Ph.D., Ami Amit,

Molecular weight determination

Presentation transcript:

Investigation of photoinduced reactivity of Glue-FITC: 1- Absorption spectra: Glue-FITC 3µM + BSA 10µM in Tris-HCl buffer (Tris – 20mM, pH- 7.0) + UV irradiation (305-315nm band pass filter) Observed decrease in the absorption intensity at 270-320 nm. Due to the photochemical covalent binding of benzophenone (BP) with protein. Glue+ BSA Glue BSA

2- Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE)

3- Kinesin/microtubule biomachinery system: + MT + MT +Glue-FITC + UV irradiation + Glue + UV irradiation Kinesin + Cy5 labeled microtubules Glue-FITC UV irradiation.

BP is essential for the covalent immobilization:

Optical tweezers: To investigate the change in the binding strength between kinesin and the microtubules upon adhesion with glue and subsequent photoclicking. Bright field microscopy traces:

Conclusion: Study demonstrated that water soluble GlueBP-FITC can serve as a photoclickable molecular glue that covalently attaches to proteins. As studied by “optical tweezers”, the temporal-to-permanent photochemical transformation gives rise to a notable change in binding strength. Protein-Glue study (with BSA and kinesin) shows that, both non covalent and covalent interaction affects the secondary structure of the proteins or their biological functions. Glue might be a potent candidate for the development of targeted cancer therapy.