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Clone
3A3
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Immunogen
Human recombinant HSP70 overexpressed in E.coli
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Antibody s full description
Mouse Anti-Human HSP70 Monoclonal IgG1 Antibody, Clone: 3A3: Unconjugated
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Antibody s category
Monoclonal Antibodies
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Antibody s other name
HSP70 1 Antibody, HSP70 2 Antibody, HSP70.1 Antibody, HSP72 Antibody, HSP73 Antibody, HSPA1 Antibody, HSPA1A Antibody, HSPA1B Antibody
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Raised in
Mouse
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Antibody s target
HSP70
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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, AM
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Antibody s reactivity
Human, Mouse, Rat, Amphibians, Chicken, Fish, Yeast, Yeast (Saccharomyces cerevisiae), Fruit Fly (Drosophila melanogaster), Bacteria, Bacteria (Shigella flexneri), Brine Shrimp (Artemia franciscanna)
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Antibody s dilutions
WB (1:5000), ICC/IF (1:500), IP (2µg); 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 pH7.2, 50% glycerol, 0.09% sodium azide
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Antibody s concentration
1 mg/ml
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Antibody s specificity
Detects ~70kDa. May detect HSP70, HSC70, p75 and HSP72.
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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
0.2 µg/ml of SMC-164 was sufficient for detection of HSP70 in 20 µg of heat shocked HeLa cell lysate by colorimetric immunoblot analysis using Goat anti-mouse IgG:HRP as the secondary antibody.
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Antibody in cell
Cytoplasm
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Tissue specificity
See included datasheet or contact our support service
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Scientific context
HSP70 genes encode abundant heat-inducible 70-kDa HSPs (HSP70s). In most eukaryotes HSP70 genes exist as part of a multigene family. They are found in most cellular compartments of eukaryotes including nuclei, mitochondria, chloroplasts, the endoplasmic reticulum and the cytosol, as well as in bacteria. The genes show a high degree of conservation, having at least 50% identity (1). The N-terminal two thirds of HSP70s are more conserved than the C-terminal third. HSP70 binds ATP with high affinity and possesses a weak ATPase activity which can be stimulated by binding to unfolded proteins and synthetic peptides (2). When HSC70 (constitutively expressed) present in mammalian cells was truncated, ATP binding activity was found to reside in an N-terminal fragment of 44 kDa which lacked peptide binding capacity. Polypeptide binding ability therefore resided within the C-terminal half (3). The structure of this ATP binding domain displays multiple features of nucleotide binding proteins (4). All HSP70s, regardless of location, bind proteins, particularly unfolded ones. The molecular chaperones of the HSP70 family recognize and bind to nascent polypeptide chains as well as partially folded intermediates of proteins preventing their aggregation and misfolding. The binding of ATP triggers a critical conformational change leading to the release of the bound substrate protein (5). The universal ability of HSP70s to undergo cycles of binding to and release from hydrophobic stretches of partially unfolded proteins determines their role in a great variety of vital intracellular functions such as protein synthesis, protein folding and oligomerization and protein transport. For more information visit our HSP70 Scientific Resource Guide at more.
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Bibliography
1. Balashova N. et al. (2005) J Biol Chem 280:2186-96. 2. Boorstein W. R., Ziegelhoffer T. & Craig E. A. (1993) J. Mol. Evol.38 (1): 1-17. 3. Rothman J. (1989) Cell 59: 591 -601. 4. DeLuca-Flaherty et al. (1990) Cell 62: 875-887. 5. Bork P., Sander C. & Valencia A. (1992) Proc. Nat Acad. Sci. USA 89: 7290-7294. 6. Fink A.L. (1999) Physiol. Rev. 79: 425-449.
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Released date
1-Nov-2008
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PubMed number
24167293
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Tested applications
Western Blot
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Tested reactivity
S. flexneri M90T
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NCBI number
NP_005336.3
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Gene number
3303
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Protein number
P08107
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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-HSP70 Monoclonal Antibody, Clone 3A3 (SMC-164). Tissue: Cervical Cancer cell line (HeLa). Species: Human. Fixation: 4% Formaldehyde for 15 min at RT. Primary Antibody: Mouse Anti-HSP70 Monoclonal Antibody (SMC-164) at 1:100 for 60 min at RT. Secondary Antibody: Goat Anti-Mouse ATTO 488 at 1:100 for 60 min at RT. Counterstain: DAPI (blue) nuclear stain at 1:5000 for 5 min RT. Localization: Cytoplasm. Magnification: 40X. Western Blot analysis of Artemia franciscanna (brine shrimp) cell lysates showing detection of Hsp70 protein using Mouse Anti-Hsp70 Monoclonal Antibody, Clone 3A3 (SMC-164). Primary Antibody: Mouse Anti-Hsp70 Monoclonal Antibody (SMC-164) at 1:1000. Courtesy of: Alison King. Western Blot analysis of Rat cell lysates showing detection of Hsp70 protein using Mouse Anti-Hsp70 Monoclonal Antibody, Clone 3A3 (SMC-164). Load: 15 µg. Block: 1.5% BSA for 30 minutes at RT. Primary Antibody: Mouse Anti-Hsp70 Monoclonal Antibody (SMC-164) at 1:1000 for 2 hours at RT. Secondary Antibody: Sheep Anti-Mouse IgG: HRP for 1 hour at RT. Mouse Anti-HSP70 Antibody [3A3] used in Immunocytochemistry/Immunofluorescence (ICC/IF) on Human Cervical Cancer cell line (HeLa) (SMC-164) Mouse Anti-Hsp70 Antibody [3A3] used in Western Blot (WB) on Artemia franciscanna (brine shrimp) cell lysates (SMC-164) Mouse Anti-Hsp70 Antibody [3A3] used in Western Blot (WB) on Rat cell lysates (SMC-164)
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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
0.025
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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