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Clone
5D12-A12
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Immunogen
Human HSP27
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Antibody s full description
Mouse Anti-Human HSP27 Monoclonal IgG2b Kappa Antibody, Clone: 5D12-A12: ATTO 390
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Antibody s category
Monoclonal Antibodies
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Antibody s other name
28kDa heat shock protein Antibody, CMT2F Antibody, HSP25 Antibody, HSP27 Antibody, HSP28 Antibody, HSPB1 Antibody, SRP27 Antibody
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Raised in
Mouse
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Antibody s target
HSP27
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Primary research fields
Cancer, Heat Shock
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Brandname
None
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Antibodies applications
WB, IHC, ICC/IF, IP, ELISA
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Antibody s reactivity
Human
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Antibody s dilutions
WB (1:2000), ICC/IF (1:100); optimal dilutions for assays should be determined by the user.
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Purity
Protein G Purified
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Antibody buffer for storage
PBS pH7.4, 50% glycerol, 0.09% sodium azide
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Antibody s concentration
1 mg/ml
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Antibody s specificity
Detects ~27kDa. Has no cross-reactivity to Alpha B crystallin. Very limited cross-reactivity to other species.
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Storage recommendations
-20ºC
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Shipping recommendations
Blue Ice or 4ºC
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Antibody certificate of analysis
0.5 µg/ml of SMC-161 was sufficient for detection of HSP27 in 10 µg of HeLa lysate by colorimetric immunoblot analysis using Goat anti-mouse IgG:HRP as the secondary antibody.
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Antibody in cell
Cytoplasm , Nucleus , Cytoskeleton , Spindle
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Tissue specificity
See included datasheet or contact our support service
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Scientific context
HSP27s belong to an abundant and ubiquitous family of small heat shock proteins (sHSP). It is an important HSP found in both normal human cells and cancer cells. The basic structure of most sHSPs is a homologous and highly conserved amino acid sequence, with an α-crystallin domain at the C-terminus and the WD/EPF domain at the less conserved N-terminus. This N-terminus is essential for the development of high molecular oligomers (1, 2). HSP27-oligomers consist of stable dimers formed by as many as 8-40 HSP27 protein monomers (3). The oligomerization status is connected with the chaperone activity: aggregates of large oligomers have high chaperone activity, whereas dimers have no chaperone activity (4). HSP27 is localized to the cytoplasm of unstressed cells but can redistribute to the nucleus in response to stress, where it may function to stabilize DNA and/or the nuclear membrane. Other functions include chaperone activity (as mentioned above), thermo tolerance in vivo, inhibition of apoptosis, and signal transduction. Specifically, in vitro, it acts as an ATP-independent chaperone by inhibiting protein aggregation and by stabilizing partially denatured proteins, which ensures refolding of the HSP70 complex. HSP27 is also involved in the apoptotic signaling pathway because it interferes with the activation of cytochrome c/Apaf-1/dATP complex, thereby inhibiting the activation of procaspase-9. It is also hypothesized that HSP27 may serve some role in cross-bridge formation between actin and myosin (5). And finally, HSP27 is also thought to be involved in the process of cell differentiation. The up-regulation of HSP27 correlates with the rate of phosphorylation and with an increase of large oligomers. It is possible that HSP27 may play a crucial role in termination of growth (6). For more information visit our HSP27 Scientific Resource Guide at more.
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Bibliography
1. Kim K.K., Kim R., and Kim, S. (1998) Nature 394(6693): 595-599. 2. Van Montfort R., Slingsby C., and Vierling E. (2001) Addv Protein Chem. 59: 105-56. 3. Ehrnsperger M., Graber S., Gaestel M. and Buchner J. (1997) EMBO J. 16: 221-229. 4. Ciocca D.R., Oesterreich S., Chamness G.C., McGuire W.L., and Fugua S.A. (1993) J Natl Cancer Inst. 85 (19): 1558-70. 5. Sarto C., Binnz P.A., and Mocarelli P. (2000) Electrophoresis. 21(6): 1218-26. 6. Arrigo A.P. (2005) J Cell Biochem. 94(2): 241-6.
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Released date
1-May-2009
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PubMed number
27909051, 24421331
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Tested applications
Western Blot, Immunohistochemistry
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Tested reactivity
Human, Human
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NCBI number
NP_001531.1
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Gene number
3315
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Protein number
P04792
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Antibody s datasheet
Contact our support service
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Representative figure link
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Representative figure legend
Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-Hsp27 Monoclonal Antibody, Clone 5D12-A3 (SMC-161). Tissue: Heat Shocked HeLa Cells. Species: Human. Fixation: 2% Formaldehyde for 20 min at RT. Primary Antibody: Mouse Anti-Hsp27 Monoclonal Antibody (SMC-161) at 1:100 for 12 hours at 4°C. Secondary Antibody: FITC Goat Anti-Mouse (green) at 1:200 for 2 hours at RT. Counterstain: DAPI (blue) nuclear stain at 1:40000 for 2 hours at RT. Localization: Cytoplasm. Nucleus. Magnification: 100x. (A) DAPI (blue) nuclear stain. (B) Anti-Hsp27 Antibody. (C) Composite. Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-Hsp27 Monoclonal Antibody, Clone 5D12-A3 (SMC-161). Tissue: HaCaT cells. Species: Human. Fixation: Cold 100% methanol for 10 minutes at -20°C. Primary Antibody: Mouse Anti-Hsp27 Monoclonal Antibody (SMC-161) at 1:100 for 1 hour at RT. Secondary Antibody: FITC Goat Anti-Mouse (green) at 1:50 for 1 hour at RT. Localization: Dull heterogeneous staining, some perinuclear, some nuclear and some cytoplasmic staining . Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-Hsp27 Monoclonal Antibody, Clone 5D12-A3 (SMC-161). Tissue: Heat Shocked HeLa Cells. Species: Human. Fixation: 2% Formaldehyde for 20 min at RT. Primary Antibody: Mouse Anti-Hsp27 Monoclonal Antibody (SMC-161) at 1:100 for 12 hours at 4°C. Secondary Antibody: FITC Goat Anti-Mouse (green) at 1:200 for 2 hours at RT. Counterstain: DAPI (blue) nuclear stain at 1:40000 for 2 hours at RT. Localization: Cytoplasm. Nucleus. Magnification: 20x. (A) DAPI (blue) nuclear stain. (B) Anti-Hsp27 Antibody. (C) Composite. Western Blot analysis of Human Cell lysates showing detection of Hsp27 protein using Mouse Anti-Hsp27 Monoclonal Antibody, Clone 5D12-A3 (SMC-161). Load: 15 µg. Block: 1.5% BSA for 30 minutes at RT. Primary Antibody: Mouse Anti-Hsp27 Monoclonal Antibody (SMC-161) at 1:1000 for 2 hours at RT. Secondary Antibody: Sheep Anti-Mouse IgG: HRP for 1 hour at RT. Mouse Anti-Hsp27 Antibody [5D12-A3] used in Immunocytochemistry/Immunofluorescence (ICC/IF) on Human Heat Shocked HeLa Cells (SMC-161) Mouse Anti-Hsp27 Antibody [5D12-A3] used in Immunocytochemistry/Immunofluorescence (ICC/IF) on Human HaCaT cells (SMC-161) Mouse Anti-Hsp27 Antibody [5D12-A3] used in Immunocytochemistry/Immunofluorescence (ICC/IF) on Human Heat Shocked HeLa Cells (SMC-161) Mouse Anti-Hsp27 Antibody [5D12-A3] used in Western Blot (WB) on Human Cell lysates (SMC-161)
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Warning information
Non-hazardous
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Country of production
Canada
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Total weight kg
1.4
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Net weight g
0.2
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Stock availability
In Stock
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Description
This antibody needs to be stored at + 4°C in a fridge short term in a concentrated dilution. Freeze thaw will destroy a percentage in every cycle and should be avoided. Antibody for research use.
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Test
StressMark antibodies supplies antibodies that are for research of human proteins. Mouse or mice from the Mus musculus species are used for production of mouse monoclonal antibodies or mabs and as research model for humans in your lab. Mouse are mature after 40 days for females and 55 days for males. The female mice are pregnant only 20 days and can give birth to 10 litters of 6-8 mice a year. Transgenic, knock-out, congenic and inbread strains are known for C57BL/6, A/J, BALB/c, SCID while the CD-1 is outbred as strain.
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Properties
Human proteins, cDNA and human recombinants are used in human reactive ELISA kits and to produce anti-human mono and polyclonal antibodies. Modern humans (Homo sapiens, primarily ssp. Homo sapiens sapiens). Depending on the epitopes used human ELISA kits can be cross reactive to many other species. Mainly analyzed are human serum, plasma, urine, saliva, human cell culture supernatants and biological samples.
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About
Monoclonals of this antigen are available in different clones. Each murine monoclonal anibody has his own affinity specific for the clone. Mouse monoclonal antibodies are purified protein A or G and can be conjugated to FITC for flow cytometry or FACS and can be of different isotypes.
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Latin name
Mus musculus