Anti-Human Hsp60 ELISA Kit
TECHNICAL SPECIFICATIONS
5777 Hine s Drive, Ann Arbor, M I 48108 USA Tol l free (800) 833 -8651 Fax (734) 668-2793 www. stressg enbioreag ents .com te chn ica l@assa yde signs. com
Anti-Human (IgG/A/M) Hsp60 ELISA Kit
Product #: EKS-650
An immunoassay for the detection and quantitation of anti-human Hsp60 antibodies in serum.
Specificity Typical Standard Curve
Detects IgG, IgA and IgM antibodies to human Hsp60.
Species Reactivity
Human
Sample Types
Serum, Plasma
Scientific Background
Heat shock protein 60 (Hsp60) is a molecular chaperone that participates in the
folding of mitochondrial proteins and facilitates proteolytic degradation of
misfolded or denatured proteins. Hsp60 is strongly dependent upon its co-
chaperone, Hsp10 that binds the molecule and regulates its substrate binding and
ATPase activity 1,2,3. Hsp60 is rapidly upregulated by a range of cellular insults
including oxidative stress, viral infection, chemical exposure, increased
References Citing EKS-650
temperature, irradiation etc. Hsp60 is mainly located in the mitochondria but
Yamazaki, K.; Ueki-Maruayama, K.; Honda, T.; Nakajima, T.; Seymour, G.J.; (2004) Clin. Exp.
changes in the intracellular location and cell surface expression have been reported
4,5,6,7,8 Immunol. 135: 478-482.
. Rev: 05/16/06
The Hsp60 family of proteins including GroEL from E. coli and Hsp65 from
mycobacteria are highly conserved during evolution and an autoimmune reaction
may be evoked because of the immunological cross-reaction between self Hsp60
CERTIFICATE OF ANALYSIS
and microbial pathogens 9,10. Human Hsp60 also shares sequence homology with a
wide range of autoantigens and the extent of this homology may contribute to
Anti-Hsp60 ELISA Kit
autoimmunity 11. Autologous Hsp60 when expressed or released from stressed
autologous cells may serve as danger signal for the innate immune system to mount
an adaptive response and both B and T cell responses to autologous Hsp60 have
Product #: EKS-650 Size: 1 X 96 well plate
been reported to occur in a variety of inflammatory conditions. Anti-Hsp60
antibody levels have been studied in the peripheral circulation of normal controls
and in the aging process 12,13.
Hsp60 has become a focus of interest as a potential autoantigen during chronic
inflammation; artherosclerosis and autoimmune diseases and anti-human Hsp60
Number of Samples: 1 standard curve and 41 samples in duplicate or 2
levels have been measured in a variety of medical conditions. Significant increases standard curves and 34 samples in duplicate
in Hsp60 autoantibody levels precede the appearance of glucose intolerance and
diabetes in cystic fibrosis patients 14. Elevated levels of circulating anti- human Sensitivity: 2.88ng/mL Range: 7.81-250ng/mL
Hsp60 antibodies are present in gastric atrophy patients and in patients with
lymphoma of mucosa associated lymphoid tissue (MALT) type which is closely
Sample Volume: 100礚/well Incubation Time: 3.25 hours
linked with host immune responses to H. pylori 15,16. Higher levels of human and
bacterial Hsp60 antibodies are present in glaucoma patients 17,18. Increases of anti-
Wavelength Setting: 450nm
Hsp60 antibodies have been reported in 43% of osteosarcoma patients 19. Levels of
anti-Hsp60 antibodies are associated with the presence and severity of coronary
Materials Included: ? Human Hsp60 Immunoassay Plate, Lot# 05BQ001
artery disease and their simultaneous presence with high levels of antibodies to
Anti-Human Hsp60 Standard, Lot# 05080605
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Chlamydia pneumoniae substantially increases the risk for disease development
Sample Diluent 1, Lot# 05A153
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20,21,22,23,24,25,26,27
. Recent data indicates that promoter polymorphism at position ?74
20X Wash Buffer, Lot# 04070618
?br>
of the IL-6 gene may be one of the genetic factors associated with high levels of
Anti-Human GAM-HRP Conjugate, Lot# 05050604
autoantibodies to Hsp60 28. Ongoing research is required to investigate the ?br>
TMB Substrate, Lot# 03170603
complexity of the relationship between circulating Hsp60 and anti-Hsp60 antibody ?br>
in the potentiation or prevention of pathogenesis. Stop Solution 2, Lot# 03150602
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Instruction Manual
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Background References
Storage of Kit: All reagents are stable as supplied at 4癈 until the kit's
References are available upon request.
expiry date.
Assay Procedure Summary The components in each kit lot # have been quality assured and
Note:
1. Add Anti-Hsp60 Standards and samples in duplicate to the pre-coated warranted in this specific combination only; please do not mix
ready-to-use r. Human Hsp60 Immunoassay Plate; incubate then wash. them with components from other kit lot #s
2. Add Anti-Human GAM-HRP Conjugate; incubate then wash.
FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC
3. Develop with TMB Substrate.
PROCEDURES: This product is not to be used for diagnostic nor
4. Add Acid Stop Solution. therapeutic purposes. By accepting this product, the user confirms that it
will be used for research purposes only. Stressgen warrants that its
5. Measure absorbance at 450nm.
products conform to the information published by Stressgen. Purchaser
6. Plot Anti-Hsp60 standard curve and determine sample concentrations. must determine the suitability of the product for their particular use.
Please refer to the MSDS for product safety information.
MATERIAL SAFETY DATA SHEET
Section 1 - Product Identification and Use
Product Name: Anti-Human (IgG/A/M) Hsp60 ELISA Kit Product# EKS-650
This product is sold only for research use by qualified laboratory personnel, and is not to be used as a drug, medical device, food additive, cosmetic,
nor household chemical. It is not to be used in diagnostic, therapeutic, consumer, agricultural, nor pesticidal applications.
Manufacturer's Name: Stressgen Biotechnologies
Street Address: 5777 Hines Drive
City, Prov. Postal Code: Ann Arbor, MI 48108 USA
Fax: (734) 668-2793
EMERGENCY PHONE: (734) 668-6113 Toll Free in North America: 1-800-833-8651
Section 2 - Hazardous Ingredients
Hazardous Ingredients:
Kit components may contain the following components:
CAS Registry Numbers Chemical Name Percentage
26172-55-4 5-chloro-2-methyl-isothiazolin-3-one <<1.15
2682-20-4 2-methyl-4isothiazolin-3-one <<0.35
3-(N-Morpholino) propanesulfonic acid
1132-61-2 N/A
54827-17-7 3,3',5,5'-tetramethylbenzidine (TMB) N/A
N/A Hydorol <0.1%
7647-14-5 Sodium Chloride N/A
7647-01-0 Hydrochloric Acid N/A
Anti-Human Hsp60 Standard: CAUTION! The toxicological properties of this material have not been fully investigated. This product
is derived from human origin. Although tests are negative for infectious diseases, this product should be handled as a potentially
infectious biological material. The serum that this product was derived from was tested by FDA approved methods as defined by the
American Red Cross. Tests were negative for HIV-1, HIV-2, HIV p24 and non-reactive to HbsAg, HVC 3 and STS. No known test
method can offer complete assurance that Hepatitis B virus, Hepatitis C virus, HIV-1, HIV-2 or other infectious agents are absent.
Section 3 - Physical Data
The physical properties of the kit components have not been investigated thoroughly.
TMB: Clear, colorless to pale yellow liquid, specific gravity 1.010
Section 4 - Fire and Explosion Hazard
Components of the kit are non-flammable. Fire and explosion data have not been investigated.
Hazardous Combustion Products:
Kit components may emit toxic vapors
TMB: carbon monoxide, carbon dioxide, nitrogen oxide fumes
Acid Stop Solution: Sulphur dioxide and sulphur trioxide fumes
Section 5 - Reactivity Data
Kit components (except TMB) are stable. Avoid contact with reducing agents, amines, strong acids, strong bases, and strong
oxidizing agents.
TMB: Unstable. Protect from direct UV light, avoid elevated temperatures, protect from moisture. Incompatible with strong
oxidizing agents, metals.
Acid Stop Solution: Stable. Avoid elevated temperatures. Incompatible with organics, alkalies, oxidizing agents.
MATERIAL SAFETY DATA SHEET
Section 6 - Toxicological Properties
The toxicological properties of the kit components have not been investigated. Kit components may burn eyes severely, be harmful if
absorbed through skin, burn skin severely, irritate respiratory tract, be harmful if inhaled, be harmful if swallowed. May cause an
allergic reaction in some individuals.
Anti-Human Hsp60 Standard: CAUTION! The toxicological properties of this material have not been fully investigated. This product
is derived from human origin. Although tests are negative for infectious diseases, this product should be handled as a potentially
infectious biological material.
TMB: Irritant to eyes and skin. May cause irritation of the digestive tract and respiratory tract.
Acid Stop Solution: Corrosive to skin and eyes. Inhalation of vapours may cause irritation of nasal and respiratory tract. Ingestion
causes gastrointestinal tract discomfort, nausea and vomiting
Section 7 - Preventative Measures
Wear chemical safety goggles and compatible chemical-resistant gloves. Avoid inhalation, contact with eyes, skin or clothing.
*****MULTIPLE COMPONENT SPILL OR LEAK PROCEDURES*****
Wear protective equipment (respirator, chemical safety goggles, rubber boots, and heavy rubber gloves).
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Ventilate area.
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Absorb on sand or vermiculite and place in closed containers for disposal.
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Dispose or mix the material with a combustible solvent and burn in a chemical incinerator equipped with an afterburner and
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scrubber.
Containers of this material may be hazardous when emptied. Emptied containers retain product residues: handle as if they were
?br>
full.
Observe all federal, state and local environmental regulations.
Section 8 - First Aid Measures
If TMB is ingested, induce vomiting and call a physician. If Stop Solution is swallowed, do not induce vomiting, rinse mouth
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with water, then drink lots of water to dilute in the stomach. For all other components, if swallowed wash out mouth with water,
provided person is conscious. Call a physician.
In case of skin contact, flush with copious amounts of water for at least 15 minutes. Remove contaminated clothing and shoes. If a
?br>
rash or other irritation develops, call a physician.
If inhaled, remove to fresh air. If breathing becomes difficult, call a physician.
?br>
In case of eye contact, flush with copious amounts of water for at least 15 minutes while separating the eyelids with fingers. Call a
?br>
physician.
Section 9 - Preparation
Prepared by: Phone#: Created: Modified:
Stressgen Biotechnologies Corp. (734) 668-6113 08/04/05 08/04/05
Toll free in North America:
1-800-833-8651
The above information is believed to be correct but does not purport to be all inclusive and shall be used only as a guide. Stressgen shall not be held liable for any
damage resulting from handling or from contact with the above product. See the Technical Specification, Packing Slip, Invoice, and Product Catalogue for additional
terms and conditions of sale.
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