Immunohistochemistry analysis of Phospho-Hsp27 [pSer82] showing staining in the cytoplasm of paraffin-embedded human skeletal muscle tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Phospho-Hsp27 [pSer82] polyclonal antibody (44534G) diluted in 3% BSA-PBS at a dilution of 1:100 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.
|Tested species reactivity||Human|
|Published species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||The antiserum was produced against a chemically synthesized phosphopeptide derived from a region of human HSP27 that contains serine 82.|
|Purification||Antigen affinity chromatography|
|Storage buffer||Dulbecco's PBS, pH 7.3, with 50% glycerol, 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/million cells|
|Immunohistochemistry (Paraffin) (IHC (P))||1:20-1:200|
|Western Blot (WB)||1-2 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.
|Western Blot (WB)||See 1 publications below|
HSP27, also known as Heat shock 27 kDa protein, is a cytoplasmic protein in interphase cells and colocalizes with mitotic spindles in mitotic cells. It translocates to the nucleus during heat shock. HSP27 belongs to the small heat shock protein (HSP20) family and is known to be an important protein in stress resistance and actin organization and associates with alpha- and beta-tubulin, microtubules and CRYAB. It also interacts with other heat shock proteins like HSPB8 and HSPBAP1. HSP27 is expressed in response to environmental stresses such as heat shock, or estrogen stimulation in MCF-7 cells. Ubiquitous expression is seen in most tissues with highest levels in heart and in tissues composed of striated and smooth muscle. Defects in HSPB1 are the cause of Charcot-Marie-Tooth disease type 2F (CMT2F) and distal hereditary motor neuronopathy type 2B (HMN2B).
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Human||Not Cited||A randomized, placebo-controlled study of the effects of the p38 MAPK inhibitor SB-681323 on blood biomarkers of inflammation in COPD patients.||Singh D,Smyth L,Borrill Z,Sweeney L,Tal-Singer R||Journal of clinical pharmacology (50:94)||2010|