-
Clone
1H11
-
Immunogen
Human native HSP70 protein
-
Antibody s full description
Mouse Anti-Human HSP70 Monoclonal IgG1 Antibody, Clone: 1H11: Biotin
-
Antibody s category
Monoclonal Antibodies
-
Antibody s other name
HSP70 1 Antibody, HSP70 2 Antibody, HSP70.1 Antibody, HSP72 Antibody, HSPA1 Antibody, HSPA1A Antibody, HSPA1B Antibody
-
Raised in
Mouse
-
Antibody s target
HSP70
-
Primary research fields
Cell Signaling, Protein Trafficking, Chaperone Proteins, Cancer, Tumor Biomarkers, Heat Shock
-
Brandname
None
-
Antibodies applications
WB, ICC/IF, FCM, FACS
-
Antibody s reactivity
Human, Mouse, Rat, Bovine, Pig
-
Antibody s dilutions
WB (1:1000), ICC/IF (1:100), FACS (1:250); optimal dilutions for assays should be determined by the user.
-
Purity
Protein G Purified
-
Antibody buffer for storage
PBS pH 7.4, 50% glycerol, 0.9% Sodium Azide
-
Antibody s concentration
1 mg/ml
-
Antibody s specificity
Binds to cell surface HSP70 without permeabilization of cell membrane in tumor cell lines. Does not bind to intracellular HSP70 at low concentrations. Detects a ~70 kDa band in Western Blot.
-
Storage recommendations
-20ºC
-
Shipping recommendations
Blue Ice or 4ºC
-
Antibody certificate of analysis
A 1:250 dilution of SMC-249 was sufficient for detection of cell surface HSP70 in HCT116 cells using Fluorescence-activated cell sorting, applied in flow cytometry, with FITC as the fluorescent probe.
-
Antibody in cell
Cell membrane
-
Tissue specificity
Induced by heat shock.
-
Scientific context
HSP70 is a highly conserved protein that is ubiquitously expressed. It can be found in chloroplasts, eukaryotic cytosol, endoplasmic reticulum, and mitochondria, but also embedded in the cell membrane and in the extracellular space (1,2,3,4). Even though HSP70 is one of the most studied heat shock proteins, the export mechanism and method of membrane insertion are not fully understood. The majority of the proteins in the heat shock family lack a consensus signal for secretion via the ER-Golgi pathway (5). Researchers have found that HSP70 may be released from cells via exosomes or secretory vesicles (6). Although HSP70 is ubiquitously expressed, there is not much information about its presence on cell surface. The finding that HSP70 is localized on the cell surface of cancer cells but not normal cells suggests a conformational change of HSP70 in the lower pH environment characteristic of cancer cells (7). The presence of cell membrane embedded HSP70 has been found to increase the stability of cancer cells, thereby protecting tumors from environmental stress (8, 9). Anti-HSP70 antibody, clone 1H11 (Catalog No. SMC-249) is unique from other commercially available antibodies in that it can bind to the extracellular region of the cell membrane embedded HSP70 protein, allowing researchers to differentiate between membrane bound and intracellular HSP70 across cancer cells types.
-
Bibliography
1. Gribaldo, S. et al. (1999) J. Bacteriol. 181, 434-443. 2. Sharma, D. & Masison, D.C. (2009) Protein Pept. Lett. 16, 571-581. 3. Sharma, D. et al. (2009) PLoS. ONE.4, e6644. 4. Kampinga, H.H. & Craig, E.A. (2010) Nat. Rev. Mol. Cell Biol. 11, 579-592. 5. Nickel, W. & Seedorf, M. (2008) Annu. Rev. Cell Dev. Biol. 24, 287-308. 6. Multhoff, G. & Hightower, L.E. (1996) Cell Stress. Chaperones. 1, 167-176. 7. De Maio, A. (2011) Cell Stress. Chaperones. 16, 235-249. 8. Horvath, I. & Vigh, L. (2010) Nature. 463, 436-438. 9. Horvath, I., Multhoff, G., Sonnleitner, A., & Vigh, L. (2008) Biochim. Biophys. Acta 1778, 1653-1664.
-
Released date
5-Oct-2016
-
PubMed number
27925208
-
Tested applications
Protein Inhibition
-
Tested reactivity
Mouse
-
NCBI number
NP_005336.3
-
Gene number
3303
-
Protein number
P0DMV8
-
Antibody s datasheet
Contact our support service
-
Representative figure link
-
Representative figure legend
Immunocytochemistry/Immunofluorescence analysis on non-permeabilized HCT116 cells using Mouse Anti-HSP70 Monoclonal Antibody, Clone 1H11: FITC conjugate (SMC-249D-FITC) showing cell membrane staining. Tissue: HCT116 cells. Species: Human. Fixation: 4% Formaldehyde. Permeabilization: None. Primary Antibody: Mouse Anti-HSP70 Monoclonal Antibody (SMC-249) at 1:100. Counterstain: Wheat germ agglutinin Texas red membrane marker; DAPI (blue) nuclear stain. Localization: Cell surface, cell membrane. (A) DAPI nuclear stain. (B) Wheat germ agglutinin Texas red. (C) HSP70 Antibody. (D) Composite. Courtesy of: Lawrence Hightower, Charles Giardina, and Didem Ozcan from University of Connecticut. Fluorescence-activated cell sorting (FACS) using Mouse Anti-HSP70 Monoclonal Antibody, Clone 1H11: FITC conjugate (SMC-249D-FITC). Tissue: Jurkat E6.1. Species: Human. Unfixed/non-permeabilized cells were incubated with (A) Anti-HSP70 control antibody, or (B) Anti-HSP70 antibody, clone 1H11 (SMC-249D-FITC), for 40 min at 4°C. Fixed/permeabilized cells were incubated with (C) Anti-HSP70 control antibody, or (D) Anti-HSP70 antibody, clone 1H11 (SMC-249D-FITC) for 40 min at 4°C. Analysis shows that Anti-HSP70 (SMC-249) can detect low levels of cell surface HSP70, and can faintly detect intracellular HSP70. Courtesy of: Dr. Elyse Ireland, Institute of Medicine, University of Chester. Western Blot analysis of Human Heat shocked HeLa cell lysates showing detection of HSP70 protein using Mouse Anti-HSP70 Monoclonal Antibody, Clone 1H11 (SMC-249). Lane 1: Molecular Weight ladder (MW). Lane 2: HeLa cell lysates. Load: 20 µg. Primary Antibody: Mouse Anti-HSP70 Monoclonal Antibody (SMC-249) at 1:1000. Immunocytochemistry/Immunofluorescence analysis on permeabilized HCT116 cells using Mouse Anti-HSP70 Monoclonal Antibody, Clone 1H11: FITC conjugate (SMC-249D-FITC) showing faint cell membrane and intracellular staining. Tissue: HCT116 cells. Species: Human. Fixation: 4% Formaldehyde. Permeabilization: 0.2% PBST. Primary Antibody: Mouse Anti-HSP70 Monoclonal Antibody (SMC-249) at 1:100. Counterstain: Wheat germ agglutinin Texas red membrane marker; DAPI (blue) nuclear stain. Localization: Cell surface, faint intracellular. (A) DAPI nuclear stain. (B) Wheat germ agglutinin Texas red. (C) HSP70 Antibody. (D) Composite. Courtesy of: Lawrence Hightower, Charles Giardina, and Didem Ozcan from University of Connecticut. Fluorescence-activated cell sorting analysis using Mouse Anti-HSP70 Monoclonal Antibody, Clone 1H11: FITC conjugate (SMC-249D-FITC). Tissue: HCT116 and HK2 cells. Species: Human. Primary Antibody: Mouse Anti-HSP70 Monoclonal Antibody (SMC-249) at 1:100 for 90 min at 4°C. Counterstain: Propidium Iodide nuclear stain. (A) HK2 cells unstained. (B) HK2 cells and HSP70 Antibody. (C) HCT116 cells unstained. (D) HCT116 cells and HSP70 Antibody. Shows that HSP70 Antibody binds to the cell surface of tumor cells but not non-tumor cells. Courtesy of: Lawrence Hightower, Charles Giardina, and Didem Ozcan from University of Connecticut. Fluorescence-activated cell sorting analysis using Mouse Anti-HSP70 Monoclonal Antibody, Clone 1H11: FITC conjugate (SMC-249D-FITC). Tissue: Jurkat E6.1, U937, MCF7 and HT29 cells . Species: Human. Fixation: No fixation. Permeabilization: None. Primary Antibody: Mouse Anti-HSP70 Monoclonal Antibody (SMC-249) at 20 µg/ml for 40 min at 4°C. Counterstain: Propidium Iodide nuclear stain at 2.5 µg/ml for 5 min at 4°C. Shows that binding of HSP70 Antibody is not cell-line specific. Courtesy of: Dr. Elyse Ireland, Institute of Medicine, University of Chester. Mouse Anti-HSP70 Antibody: FITC [1H11] used in Immunocytochemistry/Immunofluorescence (ICC/IF) on non-permeabilized Human HCT116 cells (SMC-249) Mouse Anti-HSP70 Antibody: FITC [1H11] used in Fluorescence-activated cell sorting (FACS) on Human Jurkat E6.1 cells (SMC-249) Mouse Anti-HSP70 Antibody [1H11] used in Western Blot (WB) on Human Heat shocked HeLa cell lysates (SMC-249) Mouse Anti-HSP70 Antibody: FITC [1H11] used in Immunocytochemistry/Immunofluorescence (ICC/IF) on Human HCT116 cells (SMC-249) Mouse Anti-HSP70 Antibody: FITC [1H11] used in Fluorescence-activated cell sorting (FACS) on Human HCT116 and HK2 cells (SMC-249) Mouse Anti-HSP70 Antibody: FITC [1H11] used in Fluorescence-activated cell sorting (FACS) on Human Jurkat E6.1, U937, MCF7 and HT29 cells (SMC-249)
-
Warning information
Non-hazardous
-
Country of production
Canada
-
Total weight kg
1.4
-
Net weight g
0.1
-
Stock availability
In Stock
-
-
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. For cells, cell lines and tissues in culture till half confluency. Associated membrane protein types are lipopolysaccharide selective barriers. Biological membranes include cell membranes, outer coverings of cells or organelles that allow passage of certain proteins and nuclear membranes, which cover a cell nucleus; and tissue membranes, such as mucosae and serosae.
-
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.
-
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.
-
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.
-
Latin name
Mus musculus
