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Immunofluorescent analysis of Ki67 (green) in HeLa cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were probed without (left panel) or with (right panel) a Ki67 polyclonal antibody (Product # PA5-16785) at a dilution of 1:100 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat anti-rabbit IgG secondary antibody (Product # 35552) at a dilution of 1:400 for 30 minutes at room temperature. F-Actin (red) was stained with Dylight 554 Phalloidin (Product # 21834) and nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ArrayScan or a ToxInsight Instrument at 20X magnification.
|Tested species reactivity||Human, Rat|
|Host / Isotype||Rabbit / IgG|
|Immunogen||A synthetic peptide from C-terminus of human Ki-67|
|Storage buffer||PBS, pH 7.6, with 0.2% BSA|
|Contains||15mM sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunofluorescence (IF)||1:100 - 1:500|
|Immunohistochemistry (Paraffin) (IHC (P))||1:20|
* 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.
PA5-16785 targets Ki-67 in immunohistochemistry (paraffin) and immunofluorescence applications and shows reactivity with Human and Rat samples.
The PA5-16785 immunogen is a synthetic peptide from C-terminus of human Ki-67.
This antibody was orginally validated as part of a Thermo Scientific Cellomics High Content Screening Kit. The antibody sold separately may have slightly different performance and may need to be further optimized for the best results.
Ki-67 is a nuclear protein, which is expressed in the proliferating cells. Ki-67 is preferentially expressed during late G1-, S-, M-, and G2-phases of the cell cycle, while cells in the G0 (quiescent) phase are negative for this protein.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
antigen identified by monoclonal antibody Ki-67; antigen KI-67; proliferation-related Ki-67 antigen; protein phosphatase 1, regulatory subunit 105
KIA; MIB-; MIB-1; MKI67; PPP1R105