Immunofluorescent analysis of Heat Shock Protein 90 alpha (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 Heat Shock Protein 90 alpha polyclonal antibody (Product # PA5-16341) at a dilution of 1:200 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|
|Host / Isotype||Rabbit / IgG|
|Immunogen||A synthetic 11-mer peptide, corresponding to aa 2-12 (PEETQTQDQPM-Cys) from the N-terminus end of mouse heat shock protein 90alpha (hsp90alpha)|
|Storage buffer||PBS, pH 7.4, with 0.2% BSA|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Immunofluorescence (IF)||1:100 - 1:500|
|Immunohistochemistry (Paraffin) (IHC (P))||1:50-100|
|Immunoprecipitation (IP)||10 µg/ml|
|Western Blot (WB)||5-10 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.
PA5-16341 targets Heat Shock Protein 90 alpha in immunofluorescence, immunohistochemistry (paraffin), immunoprecipitation, and western blot applications and shows reactivity with Human samples.
The PA5-16341 immunogen is a synthetic 11-mer peptide, corresponding to aa 2-12 (PEETQTQDQPM-Cys) from the N-terminus end of mouse heat shock protein 90alpha (hsp90alpha).
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.
Cells respond to heat and chemical stress by inducing the synthesis of a group of proteins referred to as heat shock proteins. HSP90 is present in cells at high levels in the cytoplasm and is composed of two separate gene products, HSP90alpha and HSP90beta.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.