Anti-Hsp60 antibody (ab46798)
Key features and details
- Rabbit polyclonal to Hsp60
- Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)
- Reacts with: Mouse, Rat, Human, Pig
- Isotype: IgG
Get better batch-to-batch reproducibility with a recombinant antibody
- Research with confidence – consistent and reproducible results with every batch
- Long-term and scalable supply – powered by recombinant technology for fast production
- Success from the first experiment – confirmed specificity through extensive validation
- Ethical standards compliant – production is animal-free
Overview
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Product name
Anti-Hsp60 antibody
See all Hsp60 primary antibodies -
Description
Rabbit polyclonal to Hsp60 -
Host species
Rabbit -
Specificity
Replenishment batches of our polyclonal antibody, ab46798 are tested in WB. Previous batches were additionally validated in Flow Cyt, ICC/IF, IHC-P and IP. These applications are still expected to work and are covered by our Abpromise guarantee. You may also be interested in our alternative recombinant antibody, ab190828.
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Tested applications
Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Mouse, Rat, Human, Pig
Predicted to work with: Chicken, Hamster, Cow, Arabidopsis thaliana, Chlamydomonas reinhardtii, Orangutan, Secale cereale, Brassica napus, Heliothis virescens, Syntrophobacter fumaroxidans, Methylacidiphilum infernorum, Thermodesulfovibrio yellowstonii, Pisum sativum -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- IHC: Human liver.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab46798 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB | (8) |
1/20000. Detects a band of approximately 60 kDa (predicted molecular weight: 60 kDa).
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IHC-P | (2) |
1/100.
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ICC/IF | (1) |
Use a concentration of 5 - 10 µg/ml.
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IP | (1) |
1/40.
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Flow Cyt (Intra) |
1/30.
ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody. |
Notes |
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WB
1/20000. Detects a band of approximately 60 kDa (predicted molecular weight: 60 kDa). |
IHC-P
1/100. |
ICC/IF
Use a concentration of 5 - 10 µg/ml. |
IP
1/40. |
Flow Cyt (Intra)
1/30. ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody. |
Target
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Function
Implicated in mitochondrial protein import and macromolecular assembly. May facilitate the correct folding of imported proteins. May also prevent misfolding and promote the refolding and proper assembly of unfolded polypeptides generated under stress conditions in the mitochondrial matrix. -
Involvement in disease
Defects in HSPD1 are a cause of spastic paraplegia autosomal dominant type 13 (SPG13) [MIM:605280]. Spastic paraplegia is a degenerative spinal cord disorder characterized by a slow, gradual, progressive weakness and spasticity of the lower limbs.
Defects in HSPD1 are the cause of leukodystrophy hypomyelinating type 4 (HLD4) [MIM:612233]; also called mitochondrial HSP60 chaperonopathy or MitCHAP-60 disease. HLD4 is a severe autosomal recessive hypomyelinating leukodystrophy. Clinically characterized by infantile-onset rotary nystagmus, progressive spastic paraplegia, neurologic regression, motor impairment, profound mental retardation. Death usually occurrs within the first two decades of life. -
Sequence similarities
Belongs to the chaperonin (HSP60) family. -
Cellular localization
Mitochondrion matrix. - Information by UniProt
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Database links
- Entrez Gene: 424059 Chicken
- Entrez Gene: 511913 Cow
- Entrez Gene: 101844216 Hamster
- Entrez Gene: 3329 Human
- Entrez Gene: 15510 Mouse
- Entrez Gene: 492279 Pig
- Entrez Gene: 63868 Rat
- Omim: 118190 Human
see all -
Alternative names
- 60 kDa chaperonin antibody
- 60 kDa heat shock protein, mitochondrial antibody
- CH60_HUMAN antibody
see all
Images
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Immunocytochemistry analysis of paraformaldehyde-fixed 0.3% Triton X permeabilized HeLa FlipIn Cell line staining with ab46798 at 1/200 dilution. Secondary antibody was Alexa fluor™ 488 goat anti-rabbit IgG H+L at 1/700 dilution. Samples were incubated with the primary antibody with blocking buffer (5% FBS in PBS) for 2 hours at 24°C. Blocking was done using 5% serum for 1 hour at 24°C.
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ab46798 staining Hsp60 in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab46798 at 1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour magenta). Nuclear DNA was labelled with DAPI (shown in blue).
Also suitable in cells fixed with 4% paraformaldehyde (10 min).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
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IHC image of Hsp60 staining in a section of formalin-fixed paraffin-embedded normal human liver* performed on a Leica Biosystems BOND® RX instrument. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab46798, 1ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre -
All lanes : Anti-Hsp60 antibody (ab46798) at 1/20000 dilution (purified)
Lane 1 : Mouse brain tissue lysate at 20 µg
Lane 2 : Mouse heart tissue lysate at 20 µg
Lane 3 : Mouse spleen tissue lysate at 20 µg
Lane 4 : Rat heart tissue lysate at 10 µg
Secondary
All lanes : HRP conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 60 kDa
Observed band size: 60 kDaBlocking Buffer: 5% NFDM/TBST
Dilution Buffer: 5% NFDM/TBST
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Immunohistochemical staining of paraffin embedded human lung adenocarcinoma with purified ab46798 at a working dilution of 1 in 100. The secondary antibody used is ab97051 Goat Anti-Rabbit IgG H&L (HRP) at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
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Overlay histogram showing HeLa cells fixed in 2% PFA and stained with purified ab46798 at a dilution of 1 in 150 (red line). The secondary antibody used was FITC goat anti-rabbit at a dilution of 1 in 150. Rabbit monoclonal IgG was used as an isotype control (black) and cells without incubation with the antibody were used as a negative control (blue line).
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ab46798 (purified) at 1/40 immunoprecipitating Hsp60 in MCF7 (Lane 1). Lane 2 - PBS. For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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Anti-Hsp60 antibody (ab46798) at 1/20000 dilution (purified) + T47-D cell lysate at 10 µg
Secondary
HRP conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 60 kDa
Observed band size: 60 kDaBlocking Buffer: 5% NFDM/TBST
Dilution Buffer: 5% NFDM/TBST
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All lanes : Anti-Hsp60 antibody (ab46798) at 1/50000 dilution (purified)
Lane 1 : MCF7 cell lysate
Lane 2 : SW480 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 60 kDa
Observed band size: 60 kDaBlocking Buffer: 5% NFDM/TBST
Dilution Buffer: 5% NFDM/TBST
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Immunohistochemical staining of paraffin embedded rat kidney with purified ab46798 at a working dilution of 1 in 100. The secondary antibody used is ab97051 Goat Anti-Rabbit IgG H&L (HRP) at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
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Immunohistochemical staining of paraffin embedded mouse cardiac muscle with purified ab46798 at a working dilution of 1 in 100. The secondary antibody used is ab97051 Goat Anti-Rabbit IgG H&L (HRP) at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
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Immunohistochemical staining of paraffin embedded human cervical carcinoma with purified ab46798 at a working dilution of 1 in 100. The secondary antibody used is ab97051 Goat Anti-Rabbit IgG H&L (HRP) at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
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Unpurified ab46798 staining Hsp60 from human colon tissue by immunohistochemistry (formalin/PFA-fixed paraffin-embedded sections). Cells were formaldehyde fixed prior to blocking in 10% serum for 2 hours at 21°C. The primary antibody was diluted 1/500 and incubated with the sample for 2 hours at 21°C. Alexa fluor® 594 goat polyclonal, diluted 1/5000, was used as the secondary.
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Immunohistochemical analysis of paraffin-embedded human breast adenocarcinoma using unpurified ab46798 at 1/250 dilution.
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Anti-Hsp60 antibody (ab46798) at 1/50000 dilution (purified) + Pig heart tissue lysate at 20 µg
Secondary
HRP conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 60 kDa
Observed band size: 60 kDaBlocking Buffer: 5% NFDM/TBST
Dilution Buffer: 5% NFDM/TBST
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Anti-Hsp60 antibody (ab46798) at 1/50000 dilution (unpurified) +
T-47D whole cell lysate (ab14899) at 10 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/2000 dilution (Goat anti -rabbit HRP)
Predicted band size: 60 kDa
Observed band size: 60 kDaSecondary antibody - anti-rabbit HRP (ab6721)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (114)
ab46798 has been referenced in 114 publications.
- Lopez-Fabuel I et al. Aberrant upregulation of the glycolytic enzyme PFKFB3 in CLN7 neuronal ceroid lipofuscinosis. Nat Commun 13:536 (2022). PubMed: 35087090
- Chambers IG et al. MRP5 and MRP9 play a concerted role in male reproduction and mitochondrial function. Proc Natl Acad Sci U S A 119:N/A (2022). PubMed: 35121660
- Salazar-Petres E et al. Placental structure, function, and mitochondrial phenotype relate to fetal size in each fetal sex in mice†. Biol Reprod 106:1292-1311 (2022). PubMed: 35293971
- Chen B et al. Pigment of Ceiba speciosa (A. St.-Hil.) Flowers: Separation, Extraction, Purification and Antioxidant Activity. Molecules 27:N/A (2022). PubMed: 35684492
- Nakano T et al. Mitochondrial surface coating with artificial lipid membrane improves the transfer efficacy. Commun Biol 5:745 (2022). PubMed: 35879398