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HeLa cells were plated on coverslips overnight. The next day cells were fixed and permeabilized using the Image-iT® Fixation/Permeabilization Kit (R37602) according to protocol. Cells were then incubated with 3 µg/mL anti-ATP synthase subunit IF1 antibody (A21355) for labeling of mitochondria for 30 min at room temperature and washed three times with dPBS. Cells were then incubated with a 1:1000 dilution of FITC-labeled secondary antibody (A16079) for 30 min and washed three times in dPBS. Cells were labeled with NucBlue® Live (R37605) and ActinRed™ 555 ReadyProbes® reagent (R37112) according to protocol. Coverslips were then mounted using ProLong® Gold antifade reagent (P36930). Images were taken using the EVOS® FL Auto Imaging System and a 40X Coverslip corrected objective (AMEP4699).
|Tested species reactivity||Mouse|
|Host / Isotype||Goat / IgG|
|Immunogen||Gamma Immunoglobins Heavy and Light chains|
|Storage buffer||PBS, pH 7.2, with 1% BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles. Store in the dark.|
|Cross Adsorption||Against bovine, goat, human, rabbit and rat IgG|
|Antibody Form||Whole Antibody|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:500-1:20,000|
* 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.
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.