Immunofluorescence analysis of HSC70 / Heat Shock Cognate Protein was performed using 70% confluent log phase Caco-2 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with HSC70 / Heat Shock Cognate Protein Mouse Monoclonal Antibody (MA3014) at 1:250 dilution in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic and membranous localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Cat, Human, Mouse, Non-human primate, Rat|
|Published species reactivity||Rat, Non-human primate, Bovine, Cat, Fish, Human, Mouse, Not Applicable|
|Host / Isotype||Mouse / IgM|
|Immunogen||Mouse spermatogenic cell protein.|
|Storage buffer||ascites diluted in PBS|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:20|
|Immunohistochemistry (Frozen) (IHC (F))||1:20-1:200|
|Western Blot (WB)||1:1,000 - 1:4,000|
* 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.
MA3-014 detects heat shock cognate protein 70 kDa (HSC70) from human, feline, primate, rat, and mouse tissues. This antibody displays slight cross-reactivity to HSP70.
MA3-014 has been successfully used in Western blot, immunohistochemistry, immunofluorescence, and immunoprecipitation procedures By Western blot, this antibody detects a 70 kDa protein representing HSC70. By 2D gel electrophoresis, MA3-014 was found to bind strongly to HSC70 and faintly to HSP70 both before and after heat shock. Immunohistochemical staining of HSC70 in mouse spermatids/spermatozoa with MA3-014 results in staining restricted to the post acrosomal region in condensing spermatids and to the midpiece in spermatozoa.
The MA3-014 antigen is mouse spermatogenic cell protein.
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.
HSC70 (also known as HSC71, HSC73, HSP73, p72, prp73) is expressed constitutively and is slightly heat-inducible. HSC70 binds to the exposed loop of clathrin light chains to promote uncoating and can also bind the cytoskeleton which may facilitate cytoskeletal rearrangements. HSC70 has been shown to stimulate lysosomal degradation of intracellular proteins and to retard both aggregation and folding of mitochondrial precursor proteins in vitro.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Two new isoforms of the human hepatoma-derived growth factor interact with components of the cytoskeleton.
MA3-014 was used in western blot to study the interaction of components of the cytoskeleton with two new isoforms of the human hepatoma-derived growth factor
|Nüße J,Mirastschijski U,Waespy M,Oetjen J,Brandes N,Rebello O,Paroni F,Kelm S,Dietz F||Biological chemistry (397:417)||2016|
Proteomic analysis of exosomes derived from human lymphoma cells.
MA3-014 was used in western blot to characterize the proteomic content of lymphoma cell-derived exosomes.
|Yao Y,Wei W,Sun J,Chen L,Deng X,Ma L,Hao S||European journal of medical research (20:null)||2015|
Expression and induction of small heat shock proteins in rat heart under chronic hyperglycemic conditions.
MA3-014 was used in western blot to examine the expression of sHsp under chronic hyperglycemic conditions in rat heart
|Reddy VS,Kumar ChU,Raghu G,Reddy GB||Archives of biochemistry and biophysics (558:1)||2014|
Proteomic analysis of exosomes secreted by human mesothelioma cells.
MA3-014 was used in western blot to conduct the proteomic profiling of exosomes secreted by human mesothelioma cells.
|Hegmans JP,Bard MP,Hemmes A,Luider TM,Kleijmeer MJ,Prins JB,Zitvogel L,Burgers SA,Hoogsteden HC,Lambrecht BN||The American journal of pathology (164:1807)||2004|
The repeated bout effect and heat shock proteins: intramuscular HSP27 and HSP70 expression following two bouts of eccentric exercise in humans.
MA3-014 was used in western blot to investigate the changes of HSP27 and HSP70 expression induced by eccentric exercise in humans
|Thompson HS,Clarkson PM,Scordilis SP||Acta physiologica Scandinavica (174:47)||2002|
Characterization and regulation of the major histocompatibility complex-encoded proteins Hsp70-Hom and Hsp70-1/2.
MA3-014 was used in western blot to study the endogenous expression and inducibility of Hsc70
|Fourie AM,Peterson PA,Yang Y||Cell stress and chaperones (6:282)||2001|
Heat shock cognate-70 gene expression declines during normal aging of the primate retina.
MA3-014 was used in western blot to characterize gene expression in the primate retinas during aging.
|Bernstein SL,Liu AM,Hansen BC,Somiari RI||Investigative ophthalmology and visual science (41:2857)||2000|
Blocking the endogenous increase in HSP 72 increases susceptibility to hypoxia and reoxygenation in isolated adult feline cardiocytes.
MA3-014 was used in western blot to investigate the consequences of inhibiting the upregulation of endogenous HSP 72 in cardiac myocytes in response to hypoxic stress
|Nakano M,Mann DL,Knowlton AA||Circulation (95:1523)||1997|
A 16-kDa protein functions as a new regulatory protein for Hsc70 molecular chaperone and is identified as a member of the Nm23/nucleoside diphosphate kinase family.
MA3-014 was used in western blot to characterize a new Nm23/nucleoside diphosphate kinase family member
|Leung SM,Hightower LE||The Journal of biological chemistry (272:2607)||1997|
Heat shock factor-1 and the heat shock cognate 70 protein associate in high molecular weight complexes in the cytoplasm of NIH-3T3 cells.
MA3-014 was used in western blot to investigate the association between HSF-1 and HSC 70 protein in the cytoplasm of NIH-3T3 cells.
|Nunes SL,Calderwood SK||Biochemical and biophysical research communications (213:1)||1995|
Protein and peptide binding and stimulation of in vitro lysosomal proteolysis by the 73-kDa heat shock cognate protein.
MA3-014 was used in western blot to investigate the functional properties of hsc73
|Terlecky SR,Chiang HL,Olson TS,Dice JF||The Journal of biological chemistry (267:9202)||1992|
A novel hsp70-like protein (P70) is present in mouse spermatogenic cells.
MA3-014 was used in western blot to investigate the distribution of hsp70-like protein in mouse germ cells
|Allen RL,O'Brien DA,Eddy EM||Molecular and cellular biology (8:828)||1988|
Response of small heat shock proteins in diabetic rat retina.
MA3-014 was used in immunohistochemistry and western blot to study the expression of small heat shock proteins in the retina of diabetic rats
|Reddy VS,Raghu G,Reddy SS,Pasupulati AK,Suryanarayana P,Reddy GB||Investigative ophthalmology and visual science (54:7674)||2013|
|Non-human primate||Not Cited||
Dissecting the role of integrin subunits alpha 2 and beta 3 in rotavirus cell entry by RNA silencing.
MA3-014 was used in flow cytometry to study the role of integrin subunits alpha2 and beta3 in rotavirus cell entry using RNA silencing
|Isa P,Sánchez-Alemán MA,López S,Arias CF||Virus research (145:251)||2009|