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Flow cytometry analysis of Chicken anti-Mouse IgG (H+L) Secondary Antibody, Alexa Fluor 647 (A21463) was performed using K-562 cells stained with alpha Tubulin Mouse Monoclonal Antibody (A11126). Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Triton™ X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with alpha Tubulin antibody or with mouse isotype control at 3-5 ug/million cells in 2.5% BSA and incubated for 2 hours at room temperature. The cells were then labeled with Chicken anti-Mouse IgG (H+L) Secondary Antibody, Alexa Fluor 647 (A21463) at a dilution of 1:500 for 1 hour at room temperature. A representative 10,000 cells were acquired and analyzed for each sample using the Attune® Acoustic Focusing Cytometer. Panel a represents cells stained with the secondary antibody alone (red histogram), isotype control (blue histogram) and alpha Tubulin Monoclonal Antibody (purple histogram). Median fluorescence intensity from the three samples is compared in panel b.
|Tested species reactivity||Mouse|
|Published species reactivity||Not Applicable|
|Host / Isotype||Chicken / IgY|
|Immunogen||Gamma Immunoglobins Heavy and Light chains|
|Conjugate||Alexa Fluor® 647|
|Storage buffer||PBS, pH 7.5|
|Contains||5mM sodium azide|
|Storage Conditions||4° C, store in dark|
|Cross Adsorption||Against human and rabbit IgG prior to conjugation|
|Antibody Form||Whole Antibody|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:500|
|Immunocytochemistry (ICC)||1-10 µg/ml|
|Immunofluorescence (IF)||1-10 µg/mL|
|Immunohistochemistry (IHC)||1-10 µg/ml|
* 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 6 publications below|
Flourescence of this long-wavelength Alexa Fluor dye is not visible by looking through a conventional fluorescence microscope.
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||DISC1 complexes with TRAK1 and Miro1 to modulate anterograde axonal mitochondrial trafficking.||Ogawa F,Malavasi EL,Crummie DK,Eykelenboom JE,Soares DC,Mackie S,Porteous DJ,Millar JK||Human molecular genetics (23:906)||2014|
|Not Applicable||Not Cited||A t(1;11) translocation linked to schizophrenia and affective disorders gives rise to aberrant chimeric DISC1 transcripts that encode structurally altered, deleterious mitochondrial proteins.||Eykelenboom JE,Briggs GJ,Bradshaw NJ,Soares DC,Ogawa F,Christie S,Malavasi EL,Makedonopoulou P,Mackie S,Malloy MP,Wear MA,Blackburn EA,Bramham J,McIntosh AM,Blackwood DH,Muir WJ,Porteous DJ,Millar JK||Human molecular genetics (21:3374)||2012|
|Not Applicable||Not Cited||The cytoplasmic domain of rhesus cytomegalovirus Rh178 interrupts translation of major histocompatibility class I leader peptide-containing proteins prior to translocation.||Richards R,Scholz I,Powers C,Skach WR,Früh K||Journal of virology (85:8766)||2011|
|Not Applicable||Not Cited||A drosophila model for amyotrophic lateral sclerosis reveals motor neuron damage by human SOD1.||Watson MR,Lagow RD,Xu K,Zhang B,Bonini NM||The Journal of biological chemistry (283:24972)||2008|
|Not Applicable||Not Cited||FAT10 plays a role in the regulation of chromosomal stability.||Ren J,Kan A,Leong SH,Ooi LL,Jeang KT,Chong SS,Kon OL,Lee CG||The Journal of biological chemistry (281:11413)||2006|
|Not Applicable||Not Cited||The hepatitis B virus X protein sensitizes HepG2 cells to UV light-induced DNA damage.||Lee AT,Ren J,Wong ET,Ban KH,Lee LA,Lee CG||The Journal of biological chemistry (280:33525)||2005|