Immunofluorescent analysis of Heat Shock Protein 27 (Hsp27, 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 with (left panel) or without (right panel) a Hsp27 monoclonal antibody (Product # MA3-015), at a dilution of 1:100 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat-anti-mouse IgG secondary antibody (Product # 35502) 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 ToxInsight at 20X magnification.
|Tested species reactivity||Dog, Human, Non-human primate, Rat|
|Published species reactivity||Rat, Human, Mouse|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Partially purified human HSP27.|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay dependent|
|Immunofluorescence (IF)||1:50 - 1:500|
|Immunohistochemistry (Paraffin) (IHC (P))||1:500|
|Immunomicroscopy (IM)||Assay Dependent|
|Immunoprecipitation (IP)||2 µl|
|Western Blot (WB)||1:500 - 1:1000|
* 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.
MA3-015 detects heat shock protein 27 kDa (HSP27) from human, rat, mouse, monkey, and canine samples.
MA3-015 has been successfully used in Western blot, immunohistochemistry (paraffin), immunoprecipitation, immunofluorescence, immunocytochemistry and ELISA procedures. By Western blot, this antibody detects an ~24 kDa protein representing HSP27 in MCF-7 cell extract. Immunohistochemical staining of human cervix with MA3-015 results in intense cytoplasmic staining of the epithelium within the parabasal cell layer. Western blot detection of HSP27 in canine heart samples is weak and may require optimization.
The MA3-015 antigen is partially purified HSP27 derived from MCF-7 cytosol. The mouse HSP27 sequence is strongly conserved within the human sequence, so cross-reactivity with mouse HSP27 is expected.
Antibodies to this protein (and modification) were previously sold as part of a Thermo Scientific Cellomics High Content Screening Kit. This replacement antibody is now recommended for researchers who need an antibody for high content cell based assays. It has been thoroughly tested and validated for cellular immunofluorescence (IF) applications. Further optimization including the selection of the most appropriate fluorescent Dylight conjugated secondary antibody may have to be performed for your high content assay.
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.
Small heat shock proteins translocate to the cytoskeleton in human skeletal muscle following eccentric exercise independently of phosphorylation.
MA3-015 was used in western blot to determine if the type of contraction affects the localization or phosphorylation of sHSPs, HSP27, and alphaB-crystallin in human muscle
|Frankenberg NT,Lamb GD,Overgaard K,Murphy RM,Vissing K||Journal of applied physiology (Bethesda, Md. : 1985) (116:1463)||2014|
Small-molecule proteostasis regulators for protein conformational diseases.
MA3-015 was used in western blot to identify small-molecule proteostasis regulators and evaluate them
|Calamini B,Silva MC,Madoux F,Hutt DM,Khanna S,Chalfant MA,Saldanha SA,Hodder P,Tait BD,Garza D,Balch WE,Morimoto RI||Nature chemical biology (8:185)||2011|
fMLP induces Hsp27 expression, attenuates NF-kappaB activation, and confers intestinal epithelial cell protection.
MA3-015 was used in western blot to study how the bacterial chemotactic peptide fMLP stimulates colonic epithelial Hsp expression under physiological conditions
|Carlson RM,Vavricka SR,Eloranta JJ,Musch MW,Arvans DL,Kles KA,Walsh-Reitz MM,Kullak-Ublick GA,Chang EB||American journal of physiology. Gastrointestinal and liver physiology (292:G1070)||2007|
Identification of 14-3-3sigma as a contributor to drug resistance in human breast cancer cells using functional proteomic analysis.
MA3-015 was used in western blot to study the role of 14-3-3sigma in drug resistance in human breast cancer cells
|Liu Y,Liu H,Han B,Zhang JT||Cancer research (66:3248)||2006|
The c-Yes 3'-UTR contains adenine/uridine-rich elements that bind AUF1 and HuR involved in mRNA decay in breast cancer cells.
MA3-015 was used in western blot to study the role of the c-Yes 3'-UTR in mRNA decay in breast cancer cells
|Sommer S,Cui Y,Brewer G,Fuqua SA||The Journal of steroid biochemistry and molecular biology (97:219)||2005|
Electromagnetic fields at mobile phone frequency induce apoptosis and inactivation of the multi-chaperone complex in human epidermoid cancer cells.
MA3-015 was used in western blot to study the induction of apoptosis and inactivation of the multi-chaperone complex in human epidermal cancer cells by electromagnetic fields at mobile phone frequency
|Caraglia M,Marra M,Mancinelli F,D'Ambrosio G,Massa R,Giordano A,Budillon A,Abbruzzese A,Bismuto E||Journal of cellular physiology (204:539)||2005|
Differential proteome analysis of replicative senescence in rat embryo fibroblasts.
MA3-015 was used in western blot to perform a deifferential proteomic analysis of replicative senescence in rat embryo fibroblasts
|Benvenuti S,Cramer R,Quinn CC,Bruce J,Zvelebil M,Corless S,Bond J,Yang A,Hockfield S,Burlingame AL,Waterfield MD,Jat PS||Molecular and cellular proteomics : MCP (1:280)||2002|
The Fas-induced apoptosis analyzed by high throughput proteome analysis.
MA3-015 was used in western blot to perform a proteomic analysis of Jurkat T-cell cytosolic proteins during Fas-induced cell death
|Gerner C,Frohwein U,Gotzmann J,Bayer E,Gelbmann D,Bursch W,Schulte-Hermann R||The Journal of biological chemistry (275:39018)||2000|
Altered expression of HSP27 and HSP70 in distal oesophageal mucosa in patients with gastro-oesophageal reflux disease subjected to fundoplication.
MA3-015 was used in immunohistochemistry to investigate the effect of heat shock protein 27 and heat shock protein 70 on mucosal defence system of the distal oesophagus
|Rantanen T,Honkanen T,Paavonen T,Rantanen L,Oksala N||European journal of surgical oncology : the journal of the European Society of Surgical Oncology and the British Association of Surgical Oncology (37:168)||2011|
Spatiotemporal expression of Hsp20 and its phosphorylation in hippocampal CA1 pyramidal neurons after transient forebrain ischemia.
MA3-015 was used in immunohistochemistry to study the effect of transient forebrain ischemia on the expression and Phosphorylation of Hsp20 in hippocampal CA1 pyramidal neurons
|Niwa M,Hara A,Taguchi A,Aoki H,Kozawa O,Mori H||Neurological research (31:721)||2009|
[The effect of resistance-related proteins on the prognosis and survival of patients with osteosarcoma: an immunohistochemical analysis].
MA3-015 was used in immunohistochemistry to examine the prognostic significance of multiple genes in osteosarcoma
|Ozger H,Eralp L,Atalar AC,Toker B,Esberk Ate¿ L,Sungur M,Bilgiç B,Ayan I||Acta orthopaedica et traumatologica turcica (43:28)||2009|
Frequent PTEN genomic alterations and activated phosphatidylinositol 3-kinase pathway in basal-like breast cancer cells.
MA3-015 was used in immunohistochemistry to study the genomic changes and pathway deregulation in basal-like breast cancer cells
|Marty B,Maire V,Gravier E,Rigaill G,Vincent-Salomon A,Kappler M,Lebigot I,Djelti F,Tourdès A,Gestraud P,Hupé P,Barillot E,Cruzalegui F,Tucker GC,Stern MH,Thiery JP,Hickman JA,Dubois T||Breast cancer research : BCR (10:null)||2009|
Expressions of HSP70 and HSP27 in hepatocellular carcinoma.
MA3-015 was used in immunohistochemistry to study the expressions of heat shock proteins in hepatocellular carcinoma and their role in carcinogenesis
|Joo M,Chi JG,Lee H||Journal of Korean medical science (20:829)||2005|
Immunohistochemical study of heat shock proteins 27, 60 and 70 in the normal human adrenal and in adrenal tumors with suppressed ACTH production.
MA3-015 was used in immunohistochemistry to investigate the expression of heat shock proteins in normal and tumoral adrenal glands
|Pignatelli D,Ferreira J,Soares P,Costa MJ,Magalhães MC||Microscopy research and technique (61:315)||2003|
Expression of heat-shock proteins hsp27, hsp70 and hsp90 in malignant epithelial tumour of the ovaries.
MA3-015 was used in immunohistochemistry to study the expression of heat-shock proteins in malignant epithelial ovarian tumors
|Elpek GO,Karaveli S,Sim¿ek T,Keles N,Aksoy NH||APMIS : acta pathologica, microbiologica, et immunologica Scandinavica (111:523)||2003|
Polyglutamine expansions cause decreased CRE-mediated transcription and early gene expression changes prior to cell death in an inducible cell model of Huntington's disease.
MA3-015 was used in immunohistochemistry to investigate the effect of polyglutamine expansions on gene expression changes in a Huntington disease model
|Wyttenbach A,Swartz J,Kita H,Thykjaer T,Carmichael J,Bradley J,Brown R,Maxwell M,Schapira A,Orntoft TF,Kato K,Rubinsztein DC||Human molecular genetics (10:1829)||2001|
Differential expression of Hsp27 in normal oesophagus, Barrett's metaplasia and oesophageal adenocarcinomas.
MA3-015 was used in immunohistochemistry to investigate the expression of heat shock protein 27 in normal and malignant cells
|Soldes OS,Kuick RD,Thompson IA,Hughes SJ,Orringer MB,Iannettoni MD,Hanash SM,Beer DG||British journal of cancer (79:595)||1999|
Heat shock proteins hsp27 and hsp70: lack of correlation with response to tamoxifen and clinical course of disease in estrogen receptor-positive metastatic breast cancer (a Southwest Oncology Group Study).
MA3-015 was used in immunohistochemistry to study whether hsp27 and hsp70 expression is correlated with clinical resistance to tamoxifen in ER-positive metastatic breast cancer
|Ciocca DR,Green S,Elledge RM,Clark GM,Pugh R,Ravdin P,Lew D,Martino S,Osborne CK||Clinical cancer research : an official journal of the American Association for Cancer Research (4:1263)||1998|
Expression of heat shock proteins in squamous cell carcinoma of the tongue: an immunohistochemical study.
MA3-015 was used in immunohistochemistry to investigate the distribution of heat shock protein proteins in squamous cell carcinoma of the tongue
|Ito T,Kawabe R,Kurasono Y,Hara M,Kitamura H,Fujita K,Kanisawa M||Journal of oral pathology and medicine : official publication of the International Association of Oral Pathologists and the American Academy of Oral Pathology (27:18)||1998|
A new marker of maturation in the cervix: the estrogen-regulated 24K protein.
MA3-015 was used in immunohistochemistry to investigate hsp27 as a new biomarker for squamous cell maturation in the cervix
|Dressler LG,Ramzy I,Sledge GW,McGuire WL||Obstetrics and gynecology (68:825)||1986|
Evidence for modulation of a 24K protein in human endometrium during the menstrual cycle.
MA3-015 was used in immunohistochemistry to investigate the expression and localization of 24K protein in the endometrium of regularly cycling women
|Ciocca DR,Asch RH,Adams DJ,McGuire WL||The Journal of clinical endocrinology and metabolism (57:496)||1983|
Regulation of human CLC-3 channels by multifunctional Ca2+/calmodulin-dependent protein kinase.
MA3-015 was used in immunocytochemistry to study the regulatory mechanism for CLC-3 channels by calcium/calmodulin-dependent protein kinase in human.
|Huang P,Liu J,Di A,Robinson NC,Musch MW,Kaetzel MA,Nelson DJ||The Journal of biological chemistry (276:20093)||2001|
Chromosomal assignments of human 27-kDa heat shock protein gene family.
MA3-015 was used in ELISA to identify the chromosomal location of hsp27
|McGuire SE,Fuqua SA,Naylor SL,Helin-Davis DA,McGuire WL||Somatic cell and molecular genetics (15:167)||1989|
Quantitative enzyme-linked immunosorbent assay for the estrogen-regulated Mr 24,000 protein in human breast tumors: correlation with estrogen and progesterone receptors.
MA3-015 was used in ELISA to investigate p24's expression level in human breast tumors and the relationship with estrogen and progesterone receptors
|Adams DJ,McGuire WL||Cancer research (45:2445)||1985|
A cDNA for the estradiol-regulated 24K protein: control of mRNA levels in MCF-7 cells.
MA3-015 was used in immunoprecipitation to characterize the estradiol-regulated 24K protein in cancer
|Moretti-Rojas I,Fuqua SA,Montgomery RA,McGuire WL||Breast cancer research and treatment (11:155)||1988|
Detection of a Mr 24,000 estrogen-regulated protein in human breast cancer by monoclonal antibodies.
MA3-015 was used in immunoprecipitation to evaluate an antibody for detecting an estrogen-regulated 24 kDa protein
|Adams DJ,Hajj H,Edwards DP,Bjercke RJ,McGuire WL||Cancer research (43:4297)||1983|