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|
|Published species reactivity||Not Applicable|
|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.
|Miscellaneous PubMed (MISC)||See 14 publications below|
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.
|Not Applicable||Not Cited||MAHRP-1, a novel Plasmodium falciparum histidine-rich protein, binds ferriprotoporphyrin IX and localizes to the Maurer's clefts.||Spycher C,Klonis N,Spielmann T,Kump E,Steiger S,Tilley L,Beck HP||The Journal of biological chemistry (278:35373)||2003|
|Not Applicable||Not Cited||Simultaneous trichromatic fluorescence detection of proteins on Western blots using an amine-reactive dye in combination with alkaline phosphatase- and horseradish peroxidase-antibody conjugates.||Martin K,Hart C,Liu J,Leung WY,Patton WF||Proteomics (3:1215)||2003|
|Not Applicable||Not Cited||Simultaneous red/green dual fluorescence detection on electroblots using BODIPY TR-X succinimidyl ester and ELF 39 phosphate.||Martin K,Hart C,Schulenberg B,Jones L,Patton WF||Proteomics (2:499)||2002|
|Not Applicable||Not Cited||Structural requirements for localization and activation of protein kinase C mu (PKC mu) at the Golgi compartment.||Hausser A,Link G,Bamberg L,Burzlaff A,Lutz S,Pfizenmaier K,Johannes FJ||The Journal of cell biology (156:65)||2002|
|Not Applicable||Not Cited||Cytosolic targeting domains of gamma and delta calmodulin-dependent protein kinase II.||Caran N,Johnson LD,Jenkins KJ,Tombes RM||The Journal of biological chemistry (276:42514)||2001|
|Not Applicable||Not Cited||Insulin-like growth factor-mediated muscle differentiation: collaboration between phosphatidylinositol 3-kinase-Akt-signaling pathways and myogenin.||Tureckova J,Wilson EM,Cappalonga JL,Rotwein P||The Journal of biological chemistry (276:39264)||2001|
|Not Applicable||Not Cited||Munc18-1 promotes large dense-core vesicle docking.||Voets T,Toonen RF,Brian EC,de Wit H,Moser T,Rettig J,Südhof TC,Neher E,Verhage M||Neuron (31:581)||2001|
|Not Applicable||Not Cited||Differential assembly of rat purinergic P2X7 receptor in immune cells of the brain and periphery.||Kim M,Spelta V,Sim J,North RA,Surprenant A||The Journal of biological chemistry (276:23262)||2001|
|Not Applicable||Not Cited||Drosophila Hsc70-4 is critical for neurotransmitter exocytosis in vivo.||Bronk P,Wenniger JJ,Dawson-Scully K,Guo X,Hong S,Atwood HL,Zinsmaier KE||Neuron (30:475)||2001|
|Not Applicable||Not Cited||Identification of the binding site for fibrinogen recognition peptide gamma 383-395 within the alpha(M)I-domain of integrin alpha(M)beta2.||Yakubenko VP,Solovjov DA,Zhang L,Yee VC,Plow EF,Ugarova TP||The Journal of biological chemistry (276:13995)||2001|
|Not Applicable||Not Cited||Yop1p, the yeast homolog of the polyposis locus protein 1, interacts with Yip1p and negatively regulates cell growth.||Calero M,Whittaker GR,Collins RN||The Journal of biological chemistry (276:12100)||2001|
|Not Applicable||Not Cited||PH-20 but not acrosin is involved in sperm penetration of the macaque zona pellucida.||Yudin AI,Vandevoort CA,Li MW,Overstreet JW||Molecular reproduction and development (53:350)||1999|
|Not Applicable||Not Cited||Colorimetric capture assay for human-immunodeficiency-virus-I reverse transcriptase activity.||Rytting AS,Akerblom L,Gronowitz JS,Källander CF||Biotechnology and applied biochemistry (29 ( Pt 3):241)||1999|
|Not Applicable||Not Cited||Vps10p cycles between the late-Golgi and prevacuolar compartments in its function as the sorting receptor for multiple yeast vacuolar hydrolases.||Cooper AA,Stevens TH||The Journal of cell biology (133:529)||1996|