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Immunodetection on a Western blot with the BOLD APB chemiluminescent substrate. Samples of protein molecular weight standards (Cat. no. P6649) containing decreasing amounts of a-tubulin were run on an SDS-polyacrylamide gel and blotted onto a PVDF membrane. After electrophoresis, the blot was stained with SYPRO® Ruby protein blot stain (Cat. no. S11791) to detect total protein. After documentation of the total protein stain (top), the blot was incubated with a mouse monoclonal anti–a-tubulin antibody (Cat. no. A11126), followed by an alkaline phosphatase conjugate of goat anti–mouse IgG antibody (Cat. no. G21060). Finally, the blot was stained with the BOLD APB chemiluminescent substrate (Cat. no. B21901) to detect the alkaline phosphatase enzyme. The chemiluminescent signal was visualized using a scanner in chemiluminescence detection mode.
|Tested species reactivity||Mouse|
|Host / Isotype||Goat / IgG|
|Immunogen||Gamma Immunoglobins Heavy and Light chains|
|Storage buffer||50mM tris, pH 8.0, with 30mM trehalose, 0.1M NaCl|
|Cross Adsorption||Against human IgG and human serum|
|Antibody Form||Whole Antibody|
|Tested Applications||Dilution *|
|ELISA (ELISA)||1:500 to 1:2,000|
|Immunohistochemistry (IHC)||1:500 to 1:2,000|
|Western Blot (WB)||1:500 to 1:2,000|
* 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.
The specific activity of the AP conjugates is approximately 150–350 units/mg. One unit is defined as the amount of enzymatic activity required to hydrolyze 1.0 micromole of p nitrophenyl phosphate to p-nitrophenol and inorganic phosphate in 60 seconds at 21°C, pH 10.
Reconstituted using 0.5 mL of deionized water to make a 2 mg/mL stock solution.
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.