Sandwich ELISA of Apolipoprotein A-1 was performed by coating wells of a 96-well plate with 100ul of an Apo A-1 rabbit oligoclonal antibody (Product # 710263) diluted to a concentration of 1ug/ml in carbonate/bicarbonate buffer (Product # 28382) overnight at 4C. Wells were blocked with 150ul of StartingBlock T20 (TBS) Blocking Buffer (Product # 37543) for 30 minutes, and 80ul of recombinant human Apo A-1 (Product # 10686-H02H-50) or recombinant mouse Apo A-1 (Product # 50918-M02H-50) was added to the plate at concentrations ranging from 1.6-1000ng/ml and incubated for 1 hour at room temperature. The plate was washed with 1X TBST (Product # 28360), and 100ul per well of an Apo A-1 mouse monoclonal antibody (Product # MA5-14670) was added to each well for 1 hour at room temperature. The plate was washed, and 100ul per well of an HRP-conjugated rabbit anti-mouse IgG cross-adsorbed secondary antibody (Product # 31452) was incubated for 30 minutes at room temperature. Detection was performed using 1-Step Ultra TMB Substrate (Product # 34028), followed by Stop Solution (Product # N600). Absorbances were read on a spectrophotometer at 450-550nm.
|Tested species reactivity||Mouse|
|Published species reactivity||Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS, pH 7.6, with 15mg/ml BSA|
|Storage Conditions||4° C|
|Cross Adsorption||Against human serum proteins|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:10,000-1:200,000|
* 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.
|Western Blot (WB)||See 1 publications below|
Concentration may vary slightly from lot-to-lot, see lot-specific datasheet for exact concentration.
Product # 31452 has been successfully used in Western blot, and ICC applications.
Antibody Specificity: This antibody reacts with the heavy chains on mouse IgG and with light chains on most mouse immunoglobulins based on immunoelectrophoresis. No antibody was detected against non-immunoglobulin serum proteins. The antibody has been tested by ELISA and/or solid-phase adsorbed to ensure minimal cross-reaction with human serum proteins. However, this antibody may cross-react with immunoglobulins from other species.
Restoration and Storage: Store product at 4°C until opened. Restore with 1.0 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 4°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-Mouse secondary antibodies are affinity-purified antibodies with well-characterized specificity for mouse 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 expression pattern of aldehyde dehydrogenase 1 (ALDH1) is an independent prognostic marker for low survival in colorectal tumors.
31452 was used in western blot to study the prognostic value of the immunohistochemical expression pattern of the cancer stem cell marker ALDH1 in colorectal cancer
|Vogler T,Kriegl L,Horst D,Engel J,Sagebiel S,Schäffauer AJ,Kirchner T,Jung A||Experimental and molecular pathology (92:111)||2012|