Immunofluorescent analysis of Goat anti-Mouse IgG (H+L) Secondary Antibody, RPE conjugate was performed using 70% confluent log phase U-2 OS cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Alpha Tubulin (DM1A) Mouse Monoclonal Antibody (62204) at 1 ug/mL (Panel a), 2.5 ug/mL (Panel b) and 5 ug/mL (Panel c) in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Secondary Antibody, RPE conjugate (M30204) a dilution of 1:400 for 45 minutes at room temperature. Nuclei were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). Panel a, b and c represent the merged images showing cytoplasmic localization. Panel d is the no primary antibody control. Panel e shows cells labeled with Arfaptin 2 Rabbit Polyclonal Antibody (402400) and Goat anti-Mouse IgG (H+L) Secondary Antibody, RPE conjugate (M30204) in order to demonstrate the specificity. The images were captured at 40X magnification.
|Tested species reactivity||Mouse|
|Host / Isotype||Goat / IgG|
|Immunogen||Gamma Immunoglobins Heavy and Light chains|
|Storage buffer||PBS with BSA|
|Contains||0.1% sodium azide|
|Storage Conditions||4° C|
|Antibody Form||Whole Antibody|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1-2.5 µg/mL|
|Immunocytochemistry (ICC)||1 to 400|
|Immunofluorescence (IF)||1 to 400|
|Western Blot (WB)||1:4000|
* 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.