Anti-HSP60 HSPD1 Monoclonal Antibody

Product Name

Anti-HSP60 HSPD1 Monoclonal Antibody

View all HSP60 Antibodies

SKU/Catalog Number

M01280-2

Size

200ug

Form

liquid

Description

Boster Bio Anti-HSP60 HSPD1 Monoclonal Antibody catalog # M01280-2. Tested in ELISA, IP, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.

Storage & Handling

Store at -20°C for one year. Avoid repeated freeze-thaw cycles.

Cite This Product

Anti-HSP60 HSPD1 Monoclonal Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # M01280-2)

Host

Mouse

Contents

PBS, 50% glycerol, 0.09% sodium azide

Clonality

Monoclonal

Clone Number

LK1

Isotype

IgG1

Immunogen

Recombinant human HSP60

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

Detects ~60kDa.

Reactive Species

M01280-2 is reactive to HSPD1 in Bovine, Chicken, Dog, Drosophila, Guinea pig, Hamster, Human, Monkey, Mouse, Pig, Plant, Rabbit, Rat, Sheep, Xenopus, Silkworm

Applications

M01280-2 is guaranteed for ELISA, Flow Cytometry, IP, IHC, WB Boster Guarantee

Observed Molecular Weight

68 kDa

Calculated molecular weight

61.055kDa

Background of HSP60

In both prokaryotic and eukaryotic cells, the misfolding and aggregation of proteins during biogenesis and under conditions of cellular stress are prevented by molecular chaperones. Members of the HSP60 family of heat shock proteins are some of the best characterized chaperones. HSP60, also known as Cpn60 or GroEl, is an abundant protein synthesized constitutively in the cell that is induced to a higher concentration after brief cell shock. It is present in many species and exhibits a remarkable sequence homology among various counterparts in bacteria, plants, and mammals with more than half of the residues identical between bacterial and mammalian HSP60 (1-3). Whereas mammalian HSP60 is localized within the mitochondria, plant HSP60, or otherwise known as Rubisco-binding protein, is located in plant chloroplasts. It has been indicated that these proteins carry out a very important biological function due to the fact that HSP60 is present in so many different species. The common characteristics of the HSP60s from the divergent species are i) high abundance, ii) induction with environmental stress such as heat shock, iii) homo-oligomeric structures of either 7 or 14 subunits which reversibly dissociate in the presence of Mg2+ and ATP, iv) ATPase activity and v) a role in folding and assembly of oligomeric protein structures (4). These similarities are supported by recent studies where the single-ring human mitochondrial homolog, HSP60 with its co-chaperonin, HSP10 were expressed in a E. coli strain, engineered so that the groE operon is under strict regulatory control. This study has demonstrated that expression of HSP60-HSP10 was able to carry out all essential in vivo functions of GroEL and its co-chaperonin, GroES (5). Another important function of HSP60 and HSP10 is their protective functions against infection and cellular stress. HSP60 has however been linked to a number of autoimmune diseases, as well as Alzheimer's, coronary artery diseases, MS, and diabetes (6-9).

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

WB (1:20000), IHC (1:100), ICC/IF (1:100), IP (1:200); optimal dilutions for assays should be determined by the user.

Validation Images & Assay Conditions

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Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-Hsp60 Monoclonal Antibody, Clone LK1, (M01280-2) . Tissue: skin Fibroblasts. Species: Human. Fixation: Cold 100% methanol for 30 minutes at -20°C . Primary Antibody: Mouse Anti-Hsp60 Monoclonal Antibody (M01280-2) at 1:1000 for senescence. Courtesy of: Valentina di Felice, University of Palermo, Italy.

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Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-Hsp60 Monoclonal Antibody, Clone LK-1 (M01280-2) . Tissue: HaCaT cells. Species: Human. Fixation: Cold 100% methanol at -20°C for 10 minutes. Primary Antibody: Mouse Anti-Hsp60 Monoclonal Antibody (M01280-2) at 1:100 for 1 hour at RT. Secondary Antibody: FITC Goat Anti-Mouse (green) at 1:50 for 1 hour at RT. Localization: Cytoplasmic Staining.

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Figure 2. IHC analysis of HSPD1 using anti-HSPD1 antibody (M01280-2).
HSPD1 was detected in paraffin-embedded section. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-HSPD1 Antibody (M01280-2) overnight at 4°C. Biotinylated goat anti Mouse IgG antibody was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1021) with DAB as the chromogen.

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Figure 3. Western blot analysis of HSPD1 using anti-HSPD1 antibody (M01280-2).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-HSPD1 antigen affinity purified polyclonal antibody (Catalog # M01280-2) at 0.5 ug/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-Mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # SA1021) with Tanon 5200 system. A specific band was detected for HSPD1.

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Figure 4. IHC analysis of HSPD1 using anti-HSPD1 antibody (M01280-2).
HSPD1 was detected in paraffin-embedded section. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-HSPD1 Antibody (M01280-2) overnight at 4°C. Biotinylated goat anti Mouse IgG antibody was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1021) with DAB as the chromogen.

Gene/Protein Information For HSPD1 (Source: Uniprot.org, NCBI)

Alternative Names

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 HSPD1 CPN60, GROEL, HLD4, HSP-60, HSP60, HSP65, HuCHA60, SPG13 heat shock protein family D (Hsp60) member 1 60 kDa heat shock protein, mitochondrial|60 kDa chaperonin|P60 lymphocyte protein|chaperonin 60|epididymis secretory sperm binding protein|heat shock 60kDa protein 1 (chaperonin)|heat shock protein 65|mitochondrial matrix protein P1|short heat shock protein 60 Hsp60s1

For more info on HSPD1, check out the HSPD1 Infographic

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In this infographic, you will see the following information for HSPD1: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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