Immunofluorescent analysis of Heat Shock Protein 27 (HSP27) (green) in HeLa cells and negative control NIH3T3 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 with HSP27 polyclonal antibody (Product # PA1-017), 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, Non-human primate, Rat|
|Published species reactivity||Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues L(10) L R G P S W D P F R C(21) of human Heat Shock Protein 27.|
|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 *|
|Immunocytochemistry (ICC)||Assay dependent|
|Immunohistochemistry (IHC)||4 µg/ml|
|Western Blot (WB)||1:1000 - 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.
|Miscellaneous PubMed (MISC)||See 1 publications below|
PA1-017 detects heat shock protein 27(hsp27) from rat, mouse, monkey, and human tissue tissues.
PA1-017 has been successfully used in Western blot, immunofluorescence, immunocytochemistry, and immunohistochemistry procedures. By Western blot, this antibody detects a ~27 kDa protein which corresponds to hsp27 from HeLa cells. Immunocytochemical staining of hsp27 in HeLa cells with this antibody results in diffuse cytoplasmic staining before heat shock and diffuse cytoplasmic along with punctate staining patterns after.
The PA1-017 immunogen is a synthetic peptide corresponding to residues L(10) L R G P S W D P F R C(21) of human Heat Shock Protein 27. This 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.
Heat shock proteins (HSP) are expressed in response to various biological stresses, including high temperatures. There are several major families of HSPs including HSP70, HSP90 and HSP100. 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 a-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.
|Not Applicable||Not Cited||
Small heat shock protein 27 (Hsp27) expression is highly induced in rat myometrium during late pregnancy and labour.
PA1-017 was used in immunohistochemistry and western blot to examine the expression of Hsp27 in rat myometrium during late pregnancy and labour
|White BG,Williams SJ,Highmore K,Macphee DJ||Reproduction (Cambridge, England) (129:115)||2005|