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Western blot analysis of Vascular Endothelial Growth Factor (VEGF) was performed by loading 1ug of indicated recombinant VEGF isoforms, and 10ul of PageRuler PlusPrestained Protein Ladder (Product # 26619) per well onto a 4-20% Tris-Glycine polyacrylamide gel. Proteins were transferred to a PVDF membrane (Product # 88518) using the G2 Fast Blotter (Product # 62288) and blocked with 5% Milk/TBST for at least 1 hour at room temperature. VEGF was detected at 14kD, 19kD and 22-30 kD using a VEGF polyclonal antibody (Product # PA5-16754) at a dilution of 1:500 in blocking buffer overnight at 4C on a rocking platform, followed by an HRP-conjugated goat anti-rabbit IgG (Fc) secondary antibody (Product # 31463) at a dilution of 1:20,000 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34076).
|Tested species reactivity||Rabbit|
|Published species reactivity||Not Applicable|
|Host / Isotype||Goat / IgG|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS, pH 7.6, with 15mg/ml BSA|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:500 - 1:5,000|
|Immunocytochemistry (ICC)||1:5,000 - 1:100,000|
|Immunofluorescence (IF)||1:5,000 - 1:100,000|
|Immunohistochemistry (IHC)||1:5,000 - 1:100,000|
|Immunoprecipitation (IP)||1:500 - 1:5,000|
|Western Blot (WB)||1:5,000 - 1:200,000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Concentration may vary slightly from lot-to-lot, see lot-specific datasheet for exact concentration.
This antibody has been successfully used in Western blot, IF, ICC, IHC, IP and FACS applications.
Antibody Specificity: This antibody reacts with the heavy chains on rabbit IgG but not with the light chains common to most rabbit immunoglobulins, based on immunoelectrophoresis. No antibody was detected against non-immunoglobulin serum proteins. However, this antibody may cross-react with immunoglobulins from other species.
Restoration and Storage: Store product at 2-8°C until opened. After opening, restore with 2.0 ml distilled water (0.8 mg/ml after restoration). Centrifuge product if it is not completely clear after standing for 1-2 hours at room temperature. To judge clarity, draw product into a pasteur pipette. Product may be stored for several weeks at -70°C as an undiluted liquid. After dilution, do not use for more than one day.
To extend the shelf-life of this product, add an equal volume of glycerol to make a final concentration of approximately 50% glycerol and store at -20°C.
Country of Origin: USA
Thermo Scientific Anti-Rabbit secondary antibodies are affinity-purified antibodies with well-characterized specificity for rabbit immunoglobulins and are useful in the detection, sorting or purification of its specified target. Secondary antibodies offer increased versatility enabling users to use many detection systems (e.g. HRP, AP, fluorescence). They can also provide greater sensitivity through signal amplification as multiple secondary antibodies can bind to a single primary antibody. Most commonly, secondary antibodies are generated by immunizing the host animal with a pooled population of immunoglobulins from the target species and can be further purified and modified (i.e. immunoaffinity chromatography, antibody fragmentation, label conjugation, etc.) to generate highly specific reagents.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
The beneficial effect of a prolyl oligopeptidase inhibitor, KYP-2047, on alpha-synuclein clearance and autophagy in A30P transgenic mouse.
31463 was used in western blot to study the role of increased autophagy in the mechanism by which a prolyl oligopeptidase inhibitor decreases accumulation of aggregated alpha-synuclein
|Savolainen MH,Richie CT,Harvey BK,Männistö PT,Maguire-Zeiss KA,Myöhänen TT||Neurobiology of disease (68:1)||2014|
|Not Applicable||Not Cited||
Reversal of mitochondrial proteomic loss in Type 1 diabetic heart with overexpression of phospholipid hydroperoxide glutathione peroxidase.
31463 was used in western blot to study the ability of overexpression of mitochondrial phospholipid PHGPx to protect the cardiac mitochondrial proteome in type 1 diabetic mice
|Baseler WA,Dabkowski ER,Jagannathan R,Thapa D,Nichols CE,Shepherd DL,Croston TL,Powell M,Razunguzwa TT,Lewis SE,Schnell DM,Hollander JM||American journal of physiology. Regulatory, integrative and comparative physiology (304:R553)||2013|
Replacement of sodium with choline in slow-cooling media improves human ovarian tissue cryopreservation.
31463 was used in immunohistochemistry to study the beneficial effects for ovarian tissue cryopreservation of substituting choline for sodium in slow-cooling media
|Talevi R,Barbato V,Mollo V,Fiorentino I,De Stefano C,Guarino FM,Gualtieri R||Reproductive biomedicine online (27:381)||2013|