Name: Anti-Hsp60 antibody [EPR18245-93]
Brand: RabMAb
SKU: ab190828
Rating: 4 (4 reviews)

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Western blot - Anti-Hsp60 antibody [EPR18245-93] (ab190828)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR18245-93] to Hsp60
Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)
Reacts with: Mouse, Rat, Human

Alexa Fluor® 488 Carrier Free

Product name

Anti-Hsp60 antibody [EPR18245-93]
See all Hsp60 primary antibodies
Description

Rabbit monoclonal [EPR18245-93] to Hsp60
Host species

Rabbit
Tested applications

Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)more details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human brain, fetal heart, fetal kidney and fetal spleen lysates; HeLa, NIH/3T3, Neuro-2a and C6 cell lysates; Mouse heart and spleen lysates; Rat brain, heart, spleen and liver lysates. IHC-P: Human, mouse and rat liver tissues. ICC/IF: HeLa and NIH/3T3 cells. Flow Cyt (intra): NIH/3T3 cells. IP: NIH/3T3 cell lysate.
General notes

Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 0.05% BSA, 40% Glycerol (glycerin, glycerine), PBS
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR18245-93
Isotype

IgG
Research areas

Alternative Versions
- Anti-Hsp60 antibody [EPR18245-93] - BSA and Azide free (ab224528)
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant human Hsp60 protein (ab113192)
Related Products

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab190828 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
WB	(4)	1/1000. Detects a band of approximately 61 kDa (predicted molecular weight: 61 kDa).
IHC-P		1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF		1/200.
IP		1/30.
Flow Cyt (Intra)		1/500.

Notes
WB 1/1000. Detects a band of approximately 61 kDa (predicted molecular weight: 61 kDa).
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/200.
IP 1/30.
Flow Cyt (Intra) 1/500.

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.
Target information above from: UniProt accession P10809 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 3329 Human
- Entrez Gene: 15510 Mouse
- Entrez Gene: 63868 Rat
- Omim: 118190 Human
- SwissProt: P10809 Human
- SwissProt: P63038 Mouse
- SwissProt: P63039 Rat
- Unigene: 595053 Human
- Unigene: 727543 Human
- Unigene: 1777 Mouse
- Unigene: 102058 Rat
see all
Alternative names
- 60 kDa chaperonin antibody
- 60 kDa heat shock protein, mitochondrial antibody
- CH60_HUMAN antibody
- Chaperonin 60 antibody
- Chaperonin, 60-KD antibody
- CPN60 antibody
- fa04a05 antibody
- GROEL antibody
- heat shock 60kDa protein 1 (chaperonin) antibody
- Heat shock protein 1 (chaperonin) antibody
- Heat shock protein 60 antibody
- Heat shock protein 65 antibody
- heat shock protein family D (Hsp60) member 1 antibody
- HLD4 antibody
- Hsp 60 antibody
- HSP 65 antibody
- HSP-60 antibody
- HSP60 antibody
- HSP65 antibody
- HSPD1 antibody
- HuCHA60 antibody
- Mitochondrial matrix protein P1 antibody
- P60 lymphocyte protein antibody
- short heat shock protein 60 Hsp60s1 antibody
- SPG13 antibody
see all

Western blot - Anti-Hsp60 antibody [EPR18245-93] (ab190828)

All lanes : Anti-Hsp60 antibody [EPR18245-93] (ab190828) at 1/1000 dilution

Lane 1 : Human brain lysate
Lane 2 : Human fetal heart lysate
Lane 3 : Human fetal kidney lysate
Lane 4 : Human fetal spleen lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution

Developed using the ECL technique.

Predicted band size: 61 kDa
Observed band size: 61 kDa

Exposure time : Lanes 1,2 and 3: 3 minutes; Lane 4:1 minute.

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-Hsp60 antibody [EPR18245-93] (ab190828)

All lanes : Anti-Hsp60 antibody [EPR18245-93] (ab190828) at 1/5000 dilution

Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate
Lane 2 : NIH/3T3 (mouse embyro fibroblast cell line) cell lysate
Lane 3 : Neuro-2a (mouse neuroblastoma cell line) cell lysate
Lane 4 : C6 (rat glial tumor cell line) cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Developed using the ECL technique.

Predicted band size: 61 kDa
Observed band size: 61 kDa

Exposure time : Lanes 1,2 and 3: 30 seconds; Lane 4:15 seconds.

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-Hsp60 antibody [EPR18245-93] (ab190828)

All lanes : Anti-Hsp60 antibody [EPR18245-93] (ab190828) at 1/1000 dilution

Lane 1 : Mouse heart lysate
Lane 2 : Mouse spleen lysate
Lane 3 : Rat brain lysate
Lane 4 : Rat heart lysate
Lane 5 : Rat spleen lysate
Lane 6 : Rat liver lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Developed using the ECL technique.

Predicted band size: 61 kDa
Observed band size: 61 kDa

Exposure time : Lane 1: 30 seconds; Lane 2: 1 minute; Lanes 3,4 and 5: 10 seconds; Lane 6: 5 seconds.

Blocking/Dilution buffer: 5% NFDM/TBST.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp60 antibody [EPR18245-93] (ab190828)

Immunohistochemical analysis of paraffin-embedded human liver tissue labeling Hsp60 with ab190828 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Granular and cytoplasmic staining on human liver (PMID: 18548335). Counter stained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp60 antibody [EPR18245-93] (ab190828)

Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling Hsp60 with ab190828 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Granular and cytoplasmic staining on mouse liver (PMID: 18548335). Counter stained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp60 antibody [EPR18245-93] (ab190828)

Immunohistochemical analysis of paraffin-embedded rat liver tissue labeling Hsp60 with ab190828 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Granular and cytoplasmic staining on rat liver (PMID: 18548335). Counter stained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-Hsp60 antibody [EPR18245-93] (ab190828)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling Hsp60 with ab190828 at 1/200 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HeLa cells.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) (red) at 1/200 dilution.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-Hsp60 antibody [EPR18245-93] (ab190828)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling Hsp60 with ab190828 at 1/200 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HeLa cells.

The nuclear counter stain is DAPI (blue). COX IV is detected with Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker (ab33985) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) preadsorbed (ab150120) (red) at 1/1000 dilution.

The negative controls are as follows:-

-ve control 1: ab190828 at 1/200 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) preadsorbed (ab150120) at 1/1000 dilution.

-ve control 2: Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker (ab33985) at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-Hsp60 antibody [EPR18245-93] (ab190828)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (mouse embyro fibroblast cell line) cells labeling Hsp60 with ab190828 at 1/200 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on NIH/3T3 cells.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) (red) at 1/200 dilution.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.
Flow Cytometry (Intracellular) - Anti-Hsp60 antibody [EPR18245-93] (ab190828)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized NIH/3T3 (mouse embyro fibroblast cell line) cell line labeling Hsp60 with ab190828 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluorr^® 488) at 1/2000 dilution was used as the secondary antibody.
Immunoprecipitation - Anti-Hsp60 antibody [EPR18245-93] (ab190828)

Hsp60 was immunoprecipitated from 0.35 mg of NIH/3T3 (mouse embyro fibroblast cell line) cell lysate with ab190828 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab190828 at 1/5000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1: NIH/3T3 cell lysate 10 µg (Input).

Lane 2: ab190828 IP in NIH/3T3 cell lysate .

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab190828 in NIH/3T3 cell lysate .

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 1 second.
Anti-Hsp60 antibody [EPR18245-93] (ab190828)

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheet download

Download

Publishing research using ab190828? Please let us know so that we can cite the reference in this datasheet.

ab190828 has been referenced in 4 publications.

Feng Y et al. Dl-3-n-Butylphthalide Alleviates Demyelination and Improves Cognitive Function by Promoting Mitochondrial Dynamics in White Matter Lesions. Front Aging Neurosci 13:632374 (2021). PubMed: 33762923
Xi H et al. FOXG1 improves mitochondrial function and promotes the progression of nasopharyngeal carcinoma. Mol Med Rep 24:N/A (2021). PubMed: 34278485
Zhang L et al. Transferrin receptor-mediated reactive oxygen species promotes ferroptosis of KGN cells via regulating NADPH oxidase 1/PTEN induced kinase 1/acyl-CoA synthetase long chain family member 4 signaling. Bioengineered 12:4983-4994 (2021). PubMed: 34369274
Zhang M et al. NAD+ repletion inhibits the endothelial-to-mesenchymal transition induced by TGF-ß in endothelial cells through improving mitochondrial unfolded protein response. Int J Biochem Cell Biol 117:105635 (2019). PubMed: 31626975

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1-4 of 4 Abreviews or Q&A

Application

Western blot

Sample

Mouse Cell lysate - whole cell (cell line)

Gel Running Conditions

Reduced Denaturing (10%)

Loading amount

130 µg

Specification

cell line

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Dec 14 2021

Application

Western blot

Sample

Mouse Cell lysate - whole cell (Leukemia T cell line)

Gel Running Conditions

Reduced Denaturing (10)

Loading amount

55 µg

Specification

Leukemia T cell line

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Dec 14 2021

Application

Western blot

Sample

Human Cell lysate - whole cell (Human lung cells)

Gel Running Conditions

Reduced Denaturing (10)

Loading amount

30 µg

Specification

Human lung cells

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Dec 08 2021

Application

Western blot

Sample

Human Cell lysate - whole cell (Lung cancer cell line)

Gel Running Conditions

Reduced Denaturing (10)

Loading amount

60 µg

Specification

Lung cancer cell line

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Dec 07 2021

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Recombinant Protein

Related Products

Applications

The Abpromise guarantee

Target

Function

Involvement in disease

Sequence similarities

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

Datasheet download

Certificate of Compliance

References (4)

Customer reviews and Q&As

Western blot abreview for Anti-Hsp60 antibody [EPR18245-93]

Western blot abreview for Anti-Hsp60 antibody [EPR18245-93]

Western blot abreview for Anti-Hsp60 antibody [EPR18245-93]

Western blot abreview for Anti-Hsp60 antibody [EPR18245-93]