Immunofluorescent analysis of Phospho-HSP27 (p-HSP27) pSer15 (green) in HeLa cells either left untreated (left panel) or treated with 10uM Anisomysin (right panel) for 30 minutes. 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 with a p-HSP27 pSer15 polyclonal antibody (Product # PA1-016), at a dilution of 1:50 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 at 20X magnification.
|Tested species reactivity||Human, Rat|
|Published species reactivity||Rabbit, Rat|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic phosphopeptide corresponding to residues L(10) L R G P (pS) W D P F R C(21) of human HSP27.|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunofluorescence (IF)||1:50 - 1:200|
|Immunoprecipitation (IP)||3 µg|
|Western Blot (WB)||1:500 - 1:2000|
* 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.
|Western Blot (WB)||See 4 publications below|
PA1-016 detects phosphorylated heat shock protein 27 (hsp27) from rat and human tissue tissues.
PA1-016 has been successfully used in Western blot, immunoprecipitation, and immunofluorescence procedures. By Western blot, this antibody detects a ~27 kDa protein which corresponds to phospho-hsp27 (Ser15) from HeLa cells. For immunolocalization, Cat. # PA1-018 is recommended.
The PA1-016 immunogen is a synthetic phosphopeptide corresponding to residues L(10) L R G P (pS) W D P F R C(21) of human HSP27. The immunizing peptide is 81% conserved in mice, rats, and hamsters. The phosphorylated peptide (Cat. # PEP-187) and non-phosphorylated peptide (Cat. # PEP-240) are available for use in neutralization and control experiments.
HSP27 also referred to as the Estrogen-Regulated 24K protein and HSP28, is one of several small heat shock proteins (HSP) produced by all organisms studied. HSP27 synthesis is induced by elevated temperature, as well as estrogen in hormone responsive cells. Interestingly, human HSP27 also shares greater than 50% homology with low molecular weight Drosophila HSP's and mammalian alpha-crystalline lens protein. Because of the estrogen responsive nature of HSP27, this protein has been studied extensively in human estrogen responsive tissues such as cervix, endometrium and breast tissue. This work has led to the suggestion that HSP27 may be a useful marker in classifying various hormone sensitive tumors.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Distension of the uterus induces HspB1 expression in rat uterine smooth muscle.
PA1-016 was used in western blot to study the expression of HspB1 in rat uterine smooth muscle
|White BG,MacPhee DJ||American journal of physiology. Regulatory, integrative and comparative physiology (301:R1418)||2011|
Phosphorylation status of heat shock protein 27 influences neurite growth in adult dorsal root ganglion sensory neurons in vitro.
PA1-016 was used in western blot to study the effect of Hsp27 phosphorylation status on neurite growth in adult dorsal root ganglion sensory neurons in vitro
|Williams KL,Mearow KM||Journal of neuroscience research (89:1160)||2011|
Possible involvement of phosphatidylinositol 3-kinase/Akt signal pathway in vasopressin-induced HSP27 phosphorylation in aortic smooth muscle A10 cells.
PA1-016 was used in western blot to study the role of the PI3K/Akt signal pathway in vasopressin-induced HSP27 phosphorylation in aortic smooth muscle cells
|Suga H,Nakajima K,Shu E,Kanno Y,Hirade K,Ishisaki A,Matsuno H,Tanabe K,Takai S,Akamatsu S,Kato K,Oiso Y,Kozawa O||Archives of biochemistry and biophysics (438:137)||2005|
Hsp27 and axonal growth in adult sensory neurons in vitro.
PA1-016 was used in western blot to demonstrate the interaction between actin and Hsp27 and its role in neurite outgrowth.
|Williams KL,Rahimtula M,Mearow KM||BMC neuroscience (6:null)||2005|