Immunofluorescent analysis of Heat Shock Protein 70 (Hsp70) in NCI-H1299 Cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with a Heat Shock Protein 70 (Hsp70) monoclonal antibody (Product # MA3-008) at a dilution of 1:100 overnight at 4 C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product # 35503). Heat Shock Protein 70 (Hsp70) staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.
|Tested species reactivity||Amphibian, Avian, Fruit fly, Fish, Human, Mouse, Rat, Yeast|
|Published species reactivity||Rat, Non-human primate, Human|
|Host / Isotype||Mouse / IgM|
|Immunogen||Human recombinant HSP70 over expressed in E. coli.|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunohistochemistry (Paraffin) (IHC (P))||1:200|
|Immunoprecipitation (IP)||2 µg|
|Western Blot (WB)||1:1000 - 1:2500|
* 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.
MA3-008 detects several members of the heat shock protein 70 kDa (HSP70) gene family including HSP70, HSC70 and, following heat shock, HSP72 from yeast, Drosophila, fish, mouse, avian, amphibian and human samples.
MA3-008 has been successfully used in Western blot, immunohistochemistry (paraffin), immunoprecipitation, and immunofluorescence procedures. By Western blot, this antibody detects proteins from ~70 kDa to ~72 kDa representing different members of the HSP70 family. 2-dimensional gel electrophoresis is required to resolve the heat induced form of these proteins from their constitutively expressed counterparts. Immunofluorescence staining of HSP70 in heat shocked HeLa cells with MA3-008 results in cytoplasmic staining.
The MA3-008 antigen is recombinant human HSP70 over-expressed in E. coli. Epitope mapping with a panel of HSP70 deletion mutants suggests that the epitope recognized is located between amino acids 437-479 of human HSP70.
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. The HSP70 family is a set of highly conserved proteins that are induced by a variety of biological stresses, including heat stress, in every organism in which the proteins have been examined. The human HSP70 family members include: HSP70, a protein which is strongly inducible in all organisms but which is also constitutively expressed in primate cells; HSP72, a 72 kDa protein that is induced exclusively under stress conditions; HSC70, or cognate protein, is a 72 kDa, constitutively expressed, protein which is involved in the uncoating of clathrin coated vesicles; GRP78, or BiP, is a glucose regulated 78 kDa protein localized in the endoplasmic reticulum; and p75, or HSP75, a 75 kDa protein that is found within the mitochondria.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Amidoxime reductase system containing cytochrome b5 type B (CYB5B) and MOSC2 is of importance for lipid synthesis in adipocyte mitochondria.
MA3-008 was used in western blot to investigate the role of the amidoxime reductase system in the lipid synthesis in adipocyte mitochondria
|Neve EP,Nordling A,Andersson TB,Hellman U,Diczfalusy U,Johansson I,Ingelman-Sundberg M||The Journal of biological chemistry (287:6307)||2012|
Heat-shock protein 70 (Hsp70) expression in four limpets of the genus Lottia: interspecific variation in constitutive and inducible synthesis correlates with in situ exposure to heat stress.
MA3-008 was used in western blot to study the inter-species variation of heat-shock protein 70 expression in the genus Lottia
|Dong Y,Miller LP,Sanders JG,Somero GN||The Biological bulletin (215:173)||2008|
Human skeletal muscle HSP70 response to training in highly trained rowers.
MA3-008 was used in western blot to study how training in rowing affects the human sekletal muscle HSP70
|Liu Y,Mayr S,Opitz-Gress A,Zeller C,Lormes W,Baur S,Lehmann M,Steinacker JM||Journal of applied physiology (Bethesda, Md. : 1985) (86:101)||1999|
Definition of extracellular localized epitopes of Hsp70 involved in an NK immune response.
MA3-008 was used in western blot to identify the extracelluar part of human Hsp70 and its role in the induction of an NK-regulated immune response
|Botzler C,Li G,Issels RD,Multhoff G||Cell stress and chaperones (3:6)||1998|
Effects of expressing human Hsp70 and its deletion derivatives on heat killing and on RNA and protein synthesis.
MA3-008 was used in western blot to study the role of heat shock protein 70 in heat response and protein synthesis
|Li L,Shen G,Li GC||Experimental cell research (217:460)||1995|
|Non-human primate||Not Cited||
Heat shock cognate protein 70 is involved in rotavirus cell entry.
MA3-008 was used in blocking/activating experiment to investigate the role of hsc70 in rotavirus cell entry
|Guerrero CA,Bouyssounade D,Zárate S,Isa P,López T,Espinosa R,Romero P,Méndez E,López S,Arias CF||Journal of virology (76:4096)||2002|