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|Antigen||Heat Shock 60kDa Protein 1 (Chaperonin) (HSPD1) Antikörper|
|Reaktivität||Human, Maus, Ratte (Rattus) Alternativen|
BioImaging (BI), Western Blotting (WB)
|5 Publikationen vorhanden|
|Hersteller||Anmelden zum Anzeigen|
ProduktdetailsAntigendetails Anwendungsinformationen Handhabung Referenzen Bilder
|Kreuzreaktivität||Maus, Ratte (Rattus)|
1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Please refer to us for technical protocols.
3. This antibody has been developed and certified for the bioimaging application. However, a routine bioimaging test is not performed on every lot. Researchers are encouraged to titrate the reagent for optimal performance.
4. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
5. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
6. Triton is a trademark of the Dow Chemical Company.
|Reinigung||The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.|
|Immunogen||Human Hsp60 Recombinant Protein|
AntigendetailsProduktdetails Anwendungsinformationen Handhabung Referenzen Bilder zurück nach oben
|Andere Bezeichnung||Hsp60 (HSPD1 Antibody Abstract)|
|Hintergrund||Heat shock proteins (Hsp) are a set of highly conserved proteins that include constitutively expressed (Hsp60, Hsp70, and Hsp90) and stress-induced (Hsp27 and Hsp72) proteins. Hsp60 is localized to the mitochondria, where it promotes mitochondrial protein folding and facilitates proteolytic degradation of misfolded or denatured proteins. It binds Hsp10, which regulates the substrate binding and ATPase activity of Hsp60. In HeLa and Jurkat mitochondria, Hsp60 associates with caspase-3 to form a complex that dissociates and releases from the mitochondria during apoptosis. In addition, Hsp60 accelerates the maturation of procaspase-3 through its ATP-dependent foldase activity. In addition to its role in protein folding, Hsp60 has also been implicated in immune function. In macrophages, its binding to the toll-like receptor-4 complex induces production of TNFalpha and nitric oxide and stimulation of a proinflammatory response. Thus, the protein folding function of Hsp60 is involved in mitochondrial protein folding in both normal and apoptotic cells, while release of Hsp60 during necrosis is thought to stimulate a proinflammatory response.|
|Pathways||Activation of Innate immune Response, Regulation of Leukocyte Mediated Immunity, Positive Regulation of Immune Effector Process, Production of Molecular Mediator of Immune Response, Positive Regulation of Endopeptidase Activity|
AnwendungsinformationenProduktdetails Antigendetails Handhabung Referenzen Bilder zurück nach oben
1. Seed the cells in appropriate culture medium at ~10,000 cells per well in an 96-well Imaging Plate and culture overnight.
2. Remove the culture medium from the wells, and fix the cells by adding 100 myl of Fixation Buffer to each well. Incubate for 10 minutes at room temperature (RT).
3. Remove the fixative from the wells, and permeabilize the cells using either 90% methaol, or Triton™ X-100: a. Add 100 myl of -20°C 90% methanol to each well and incubate for 5 minutes at RT. OR b. Add 100 myl of 0.1% Triton™ X-100 to each well and incubate for 5 minutes at RT.
4. Remove the permeabilization buffer, and wash the wells twice with 100 myl of 1× PBS.
5. Remove the PBS, and block the cells by adding 100 myl of to each well. Incubate for 30 minutes at RT.
6. Remove the blocking buffer and add 50 myl of the optimally titrated primary antibody (diluted in Stain Buffer) to each well, and incubate for 1 hour at RT.
7. Remove the primary antibody, and wash the wells three times with 100 myl of 1× PBS.
8. Remove the PBS, and add the second step reagent at its optimally titrated concentration in 50 myl to each well, and incubate in the dark for 1 hour at RT.
9. Remove the second step reagent, and wash the wells three times with 100 myl of 1× PBS.
10. Remove the PBS, and counter-stain the nuclei by adding 200 myl per well of 2 myg/ml Hoechst 33342 in 1× PBS to each well at least 15 minutes before imaging.
11. View and analyze the cells on an appropriate imaging instrument.
Related Products: ABIN967389, ABIN968537
|Beschränkungen||Nur für Forschungszwecke einsetzbar|
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|Buffer||Aqueous buffered solution containing BSA, glycerol, and ≤0.09 % sodium azide.|
|Vorsichtsmaßnahmen||This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.|
|Informationen zur Lagerung||Store undiluted at -20°C.|
ReferenzenProduktdetails Antigendetails Anwendungsinformationen Handhabung Bilder zurück nach oben
|Produkt verwendet in:||
Ohashi, Burkart, Flohé, Kolb: "Cutting edge: heat shock protein 60 is a putative endogenous ligand of the toll-like receptor-4 complex." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 164, Issue 2, pp. 558-61, 2000
Samali, Cai, Zhivotovsky, Jones, Orrenius: "Presence of a pre-apoptotic complex of pro-caspase-3, Hsp60 and Hsp10 in the mitochondrial fraction of jurkat cells." in: The EMBO journal, Vol. 18, Issue 8, pp. 2040-8, 1999
Xanthoudakis, Roy, Rasper, Hennessey, Aubin, Cassady, Tawa, Ruel, Rosen, Nicholson: "Hsp60 accelerates the maturation of pro-caspase-3 by upstream activator proteases during apoptosis." in: The EMBO journal, Vol. 18, Issue 8, pp. 2049-56, 1999
Venner, Singh, Gupta: "Nucleotide sequences and novel structural features of human and Chinese hamster hsp60 (chaperonin) gene families." in: DNA and cell biology, Vol. 9, Issue 8, pp. 545-52, 1991
Gutiérrez, Rojas, Lomonte, Gené, Chaves, Alvarado, Rojas: "Standardization of assays for testing the neutralizing ability of antivenoms." in: Toxicon : official journal of the International Society on Toxinology, Vol. 28, Issue 10, pp. 1127-9; author reply 1129-32, 1991
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|Bilder des Herstellers||