HSPD1 antibody

Catalog No. ABIN968591

Product Details anti-HSPD1 Antibody

Target: HSPD1 (Heat Shock 60kDa Protein 1 (Chaperonin) (HSPD1))

Reactivity: Human, Mouse, Rat

Host: Mouse

Clonality: Monoclonal

Conjugate: This HSPD1 antibody is un-conjugated

Application: Western Blotting (WB), BioImaging (BI)

Cross-Reactivity: Mouse (Murine), Rat (Rattus)

Characteristics: 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.

Purification: The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Immunogen: Human Hsp60 Recombinant Protein

Clone: 24-HSP60

Isotype: IgG1

Application Details

Application Notes: Bioimaging
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.

Comment:

Related Products: ABIN967389, ABIN968537

Restrictions: For Research Use only

Handling

Format: Liquid

Concentration: 250 μg/mL

Buffer: Aqueous buffered solution containing BSA, glycerol, and ≤0.09 % sodium azide.

Preservative: Sodium azide

Precaution of Use: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Storage: -20 °C

Storage Comment: Store undiluted at -20°C.

References for anti-HSPD1 antibody (ABIN968591)

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) (PubMed).

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) (PubMed).

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) (PubMed).

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) (PubMed).

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) (PubMed).

Target Details for HSPD1

Target: HSPD1 (Heat Shock 60kDa Protein 1 (Chaperonin) (HSPD1))

Alternative Name: Hsp60 (HSPD1 Products)

Synonyms: CPN60 antibody, GROEL antibody, HLD4 antibody, HSP-60 antibody, HSP60 antibody, HSP65 antibody, HuCHA60 antibody, SPG13 antibody, chaperonin antibody, cpn60 antibody, groel antibody, hld4 antibody, hsp60 antibody, hsp65 antibody, spg13 antibody, 60kDa antibody, Hsp60 antibody, 12 antibody, BP5 antibody, CG12101 antibody, Cpn60 antibody, Dmel\\CG12101 antibody, Dmhsp60 antibody, G62 antibody, HSP60A antibody, Hsp60A antibody, IEF16 antibody, Mmp-P1 antibody, SSP 7506 antibody, d-hsp60 antibody, hsp60A antibody, l(1)10Ac antibody, l(1)BP5 antibody, l(1)G8 antibody, l(1)HM21 antibody, l(1)L12 antibody, l(1)dp025 antibody, cb863 antibody, fa04a05 antibody, fb22d10 antibody, fi27b05 antibody, id:ibd2197 antibody, sb:cb144 antibody, wu:fa04a05 antibody, wu:fb22d10 antibody, wu:fi04a12 antibody, wu:fi27b05 antibody, MIF4 antibody, MNA2 antibody, mopA antibody, groL antibody, crpA antibody, Hspd1-30p antibody, heat shock protein family D (Hsp60) member 1 antibody, heat shock protein family D (Hsp60) member 1 S homeolog antibody, 60 kDa heat shock protein, mitochondrial antibody, heat shock protein 1 (chaperonin) antibody, Heat shock protein 60A antibody, heat shock 60 protein 1 antibody, chaperone ATPase HSP60 antibody, molecular chaperone GroEL antibody, thermosome subunit antibody, chaperonin GroEL antibody, mitochondrial chaperonin antibody, heat shock protein family D member 1 antibody, HSPD1 antibody, hspd1.S antibody, hspd1 antibody, LOC100414401 antibody, Hspd1 antibody, Hsp60A antibody, HSP60 antibody, groEL antibody, MMP_RS07785 antibody, groEl antibody, LOC100136430 antibody

Background: 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.

Molecular Weight: 60 kDa

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