HSP60 Antibody (264233) [Unconjugated]

Western blot shows lysates of MCF-7 human breast cancer cell line, DA3 mouse myeloma cell line, and Nb2-11 rat lymphoma cell line. PVDF membrane was probed with 0.5 µg/mL of Mouse Anti-Human/Mouse/Rat HSP60 Monoclonal Antibody (Catalog # MAB1800) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for HSP60 at approximately 62 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.

HSP60 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Mouse Anti-Human/Mouse/Rat HSP60 Monoclonal Antibody (Catalog # MAB1800) at 25 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Simple Western lane view shows lysates of Jurkat human acute T cell leukemia cell line, HeLa human cervical epithelial carcinoma cell line, C2C12 mouse myoblast cell line, and C6 rat glioma cell line, loaded at 0.2 mg/mL. A specific band was detected for HSP60 at approximately 60 kDa (as indicated) using 1 µg/mL of Mouse Anti-Human/Mouse/Rat HSP60 Monoclonal Antibody (Catalog # MAB1800) . This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Hsp60 induction and release after MPP+ treatment. (A) Hsp60 immunoblot analysis and quantification in cell lysates from SH-SY5Y after treatment with 100 μM MPP+ (at 6, 12, 24 and 48 hours) compared to control cells. (B) Hsp60 released by cultured SH-SY5Y cells treated by MPP+ (100 μM, at 6, 12, 24 and 48 hours) compared to control cultures. (C) MTT and lactate dehydrogenase (LDH) assay in SH-SY5Y cells exposed to MPP+ (100 μM, at 6, 12, 24 and 48 hours) compared to control cells. MTT assay (black bars), LDH assay (grey bars). *P < 0.05 versus corresponding controls. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/24886419), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu, Mu, Rt
• Applications: WB, Simple Western, ICC/IF
• Clone: 264233
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: LYOPH

HSP60 Antibody (264233) [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant human HSP60
  Met1-Phe573
  Accession # P10809
• Specificity: Detects human, mouse and rat HSP60.
• Source: N/A
• Isotype: IgG2b
• Clonality: Monoclonal
• Host: Mouse
• Gene: HSPD1
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 8-25 ug/mL
  • Simple Western 1 ug/mL
  • Western Blot 0.5 ug/mL
• Application Notes: In Simple Western only 10-15 uL of the recommended dilution is used per data point.

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for HSP60 Antibody (264233) [Unconjugated]

• 60 kDa chaperonin
• Chaperonin 60
• cpn60
• GROEL
• heat shock 60kD protein 1 (chaperonin)
• heat shock 60kDa protein 1 (chaperonin)
• Heat shock protein 60
• heat shock protein 65
• HLD4
• HSP60
• HSP-60
• HSP60SPG13
• HSP65
• HSPD1
• HuCHA60
• Mitochondrial matrix protein P1,60 kDa heat shock protein, mitochondrial
• P60 lymphocyte protein
• short heat shock protein 60 Hsp60s1
• spastic paraplegia 13 (autosomal dominant)
• SPG13

Background

Heat shock proteins (HSPs) are a family of highly conserved stress response proteins. Heat shock proteins function primarily as molecular chaperones by facilitating the folding of other cellular proteins, preventing protein aggregation or targeting improperly folded proteins to specific degradative pathways. HSPs are typically expressed at low levels under normal physiological conditions but are dramatically upregulated in response to cellular stress. Heat Shock Protein 60 (HSP60), also known as Chaperonin 60 (CPN60), is a mitochondrial matrix protein belonging to a highly conserved family of molecular chaperone and stress response proteins. HSP60 plays a role in stabilizing and refolding proteins in response to heat-shock or other cellular stress. Full length human HSP60 is 98% identical to mouse and rat HSP60.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for HSP60 Antibody (MAB1800)(8)

Publications using MAB1800

• KT Stein, SJ Moon, AN Nguyen, HD Sikes Kinetic modeling of H2O2 dynamics in the mitochondria of HeLa cells PLoS Comput. Biol., 2020-09-14;16(9):e1008202. 2020-09-14 [PMID: 32925922] (Western Blot, Human)
• SY Choi, R Lopez-Gonz, G Krishnan, HL Phillips, AN Li, WW Seeley, WD Yao, S Almeida, FB Gao C9ORF72-ALS/FTD-associated poly(GR) binds Atp5a1 and compromises mitochondrial function in vivo Nat. Neurosci., 2019-05-13;0(0):. 2019-05-13 [PMID: 31086314] (IHC-Fr, Mouse)
• J Wettmarsha, V Goh, KT Huang, DM Arduino, U Tripathi, A Leimpek, Y Cheng, AA Pittis, T Gabaldón, D Mokranjac, G Hajnóczky, F Perocchi MICU1 Confers Protection from MCU-Dependent Manganese Toxicity Cell Rep, 2018-11-06;25(6):1425-1435.e7. 2018-11-06 [PMID: 30403999] (Western Blot, Human)
• Kassi T. Stein, Sun Jin Moon, Hadley D. Sikes Mitochondrial H2O2 Generation Using a Tunable Chemogenetic Tool To Perturb Redox Homeostasis in Human Cells and Induce Cell Death ACS Synthetic Biology 2018-09-21 [PMID: 30138563] (Western Blot, Human)
• Guangxin Li Comparative analysis of the interaction of HSPs in dendritic cells, macrophages, RGM-1 cells infected by Helicobacter pylori Am J Transl Res, 2016-10-15;8(10):4184-4194. 2016-10-15 [PMID: 27830002] (Western Blot, Rat)
• Noelker C, Morel L, Osterloh A, Alvarez-Fischer D, Lescot T, Breloer M, Gold M, Oertel W, Henze C, Michel P, Dodel R, Lu L, Hirsch E, Hunot S, Hartmann A Heat shock protein 60: an endogenous inducer of dopaminergic cell death in Parkinson disease. J Neuroinflammation, 2014-05-08;11(0):86. 2014-05-08 [PMID: 24886419] (Western Blot, Human)
• Jing Ji, Yulia Y. Tyurina, Minke Tang, Weihong Feng, Donna B. Stolz, Robert S.B. Clark et al. Mitochondrial injury after mechanical stretch of cortical neurons in vitro: biomarkers of apoptosis and selective peroxidation of anionic phospholipids Journal of Neurotrauma 3/20/2012 [PMID: 21895519]
• Gotoh T, Oyadomari S, Mori K, Mori M Nitric oxide-induced apoptosis in RAW 264.7 macrophages is mediated by endoplasmic reticulum stress pathway involving ATF6 and CHOP. J. Biol. Chem., 2002-01-22;277(14):12343-50. 2002-01-22 [PMID: 11805088] (Western Blot, Human)

Bioinformatics

• Gene Symbol: HSPD1
• Uniprot:
  • P10809(Mouse)