Anti-Hsp60 antibody (ab46798) is a rabbit polyclonal antibody that is used to detect Hsp60 in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IP, IHC-P, ICC/IF. Suitable for Human, Mouse, Pig, Rat samples.
- Over 130 publications
- Trusted since 2007
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- Science advances 9:eadi24652023Inhibition of ACLY overcomes cancer immunotherapy resistance via polyunsaturated fatty acids peroxidation and cGAS-STING activation.Applications:Unspecified applicationReactive species:Unspecified reactive speciesWei Xiang,Hongwei Lv,Fuxue Xing,Xiaoyan Sun,Yue Ma,Lu Wu,Guishuai Lv,Qianni Zong,Liang Wang,Zixin Wu,Qiyu Feng,Wen Yang,Hongyang WangPubMed 38055816
- Communications biology 6:7872023Identification of genotype-biochemical phenotype correlations associated with fructose 1,6-bisphosphatase deficiency.Applications:Unspecified applicationReactive species:Unspecified reactive speciesIkki Sakuma,Hidekazu Nagano,Naoko Hashimoto,Masanori Fujimoto,Akitoshi Nakayama,Takahiro Fuchigami,Yuki Taki,Tatsuma Matsuda,Hiroyuki Akamine,Satomi Kono,Takashi Kono,Masataka Yokoyama,Motoi Nishimura,Koutaro Yokote,Tatsuki Ogasawara,Yoichi Fujii,Seishi Ogawa,Eunyoung Lee,Takashi Miki,Tomoaki TanakaPubMed 37507476
- Nature metabolism 5:1290-13022023Fatty acid oxidation organizes mitochondrial supercomplexes to sustain astrocytic ROS and cognition.Applications:Unspecified applicationReactive species:Unspecified reactive speciesBrenda Morant-Ferrando,Daniel Jimenez-Blasco,Paula Alonso-Batan,Jesús Agulla,Rebeca Lapresa,Dario Garcia-Rodriguez,Sara Yunta-Sanchez,Irene Lopez-Fabuel,Emilio Fernandez,Peter Carmeliet,Angeles Almeida,Marina Garcia-Macia,Juan P BolañosPubMed 37460843
- Nature 618:849-8542023A cytosolic surveillance mechanism activates the mitochondrial UPR.Applications:Unspecified applicationReactive species:Unspecified reactive speciesF X Reymond Sutandy,Ines Gößner,Georg Tascher,Christian MünchPubMed 37286597
- Life (Basel, Switzerland) 13:2023Aerobic Exercise Delays Alzheimer's Disease by Regulating Mitochondrial Proteostasis in the Cerebral Cortex and Hippocampus.Applications:Unspecified applicationReactive species:Unspecified reactive speciesKaiyin Cui,Chaoyang Li,Guoliang FangPubMed 37240849
- Cancers 15:2023Immunomorphological Patterns of Chaperone System Components in Rare Thyroid Tumors with Promise as Biomarkers for Differential Diagnosis and Providing Clues on Molecular Mechanisms of Carcinogenesis.Applications:Unspecified applicationReactive species:Unspecified reactive speciesLetizia Paladino,Radha Santonocito,Giuseppa Graceffa,Calogero Cipolla,Alessandro Pitruzzella,Daniela Cabibi,Francesco Cappello,Everly Conway de Macario,Alberto J L Macario,Fabio Bucchieri,Francesca RappaPubMed 37190332
- Nature metabolism 5:642-6592023Formate overflow drives toxic folate trapping in MTHFD1 inhibited cancer cells.Applications:Unspecified applicationReactive species:Unspecified reactive speciesAlanna C Green,Petra Marttila,Nicole Kiweler,Christina Chalkiadaki,Elisée Wiita,Victoria Cookson,Antoine Lesur,Kim Eiden,François Bernardin,Karl S A Vallin,Sanjay Borhade,Maeve Long,Elahe Kamali Ghahe,Julio J Jiménez-Alonso,Ann-Sofie Jemth,Olga Loseva,Oliver Mortusewicz,Marianne Meyers,Elodie Viry,Annika I Johansson,Ondřej Hodek,Evert Homan,Nadilly Bonagas,Louise Ramos,Lars Sandberg,Morten Frödin,Etienne Moussay,Ana Slipicevic,Elisabeth Letellier,Jérôme Paggetti,Claus Storgaard Sørensen,Thomas Helleday,Martin Henriksson,Johannes MeiserPubMed 37012496
- Cell reports 41:1117892022The mitochondrial UPR regulator ATF5 promotes intestinal barrier function via control of the satiety response.Applications:Unspecified applicationReactive species:Unspecified reactive speciesDouja Chamseddine,Siraje A Mahmud,Aundrea K Westfall,Todd A Castoe,Rance E Berg,Mark W PellegrinoPubMed 36516750
- Cancer science 114:1056-10662022Mitochondrial DNA haplogroup M7: A predictor of poor prognosis for colorectal cancer patients in Chinese population.Applications:Unspecified applicationReactive species:Unspecified reactive speciesZeyu Yan,Qing Yuan,Yiwei He,Fan Peng,Yang Liu,Huanqin Zhang,Xiaoying Ji,Xianli He,Qi Zhao,Jinliang Xing,Xu GuoPubMed 36382493
- eLife 11:2022Two neuronal peptides encoded from a single transcript regulate mitochondrial complex III in .Applications:Unspecified applicationReactive species:Unspecified reactive speciesJustin A Bosch,Berrak Ugur,Israel Pichardo-Casas,Jordan Rabasco,Felipe Escobedo,Zhongyuan Zuo,Ben Brown,Susan Celniker,David A Sinclair,Hugo J Bellen,Norbert PerrimonPubMed 36346220
Key facts
- Isotype
- IgG
- Host species
- Rabbit
- Storage buffer
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA- Form
- Liquid
- Clonality
- Polyclonal
Immunogen
- The exact immunogen used to generate this antibody is proprietary information.
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Reactivity data
Select an application| IP | Flow Cyt (Intra) | WB | IHC-P | ICC/IF | |
|---|---|---|---|---|---|
| Human | Tested | Tested | Tested | Tested | Tested |
| Mouse | Expected | Expected | Tested | Tested | Expected |
| Rat | Expected | Expected | Expected | Tested | Expected |
| Chicken | Predicted | Predicted | Predicted | Predicted | Predicted |
| Cow | Predicted | Predicted | Predicted | Predicted | Predicted |
| Hamster | Predicted | Predicted | Predicted | Predicted | Predicted |
| Orangutan | Predicted | Predicted | Predicted | Predicted | Predicted |
| Pig | Expected | Expected | Tested | Expected | Expected |
IP
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info 1/40 | Notes - |
ExpectedExpected
| Species | Dilution info | Notes |
|---|---|---|
Species Mouse, Pig, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
PredictedPredicted
| Species | Dilution info | Notes |
|---|---|---|
Species Chicken, Hamster, Cow, Orangutan | Dilution info - | Notes - |
Flow Cyt (Intra)
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info 1/30 | Notes ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody. |
ExpectedExpected
| Species | Dilution info | Notes |
|---|---|---|
Species Mouse, Pig, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
PredictedPredicted
| Species | Dilution info | Notes |
|---|---|---|
Species Chicken, Hamster, Cow, Orangutan | Dilution info - | Notes - |
WB
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Mouse | Dilution info 1/20000 | Notes - |
Species Pig | Dilution info 1/20000 | Notes - |
Species Human | Dilution info 1/20000 | Notes - |
ExpectedExpected
| Species | Dilution info | Notes |
|---|---|---|
Species Rat | Dilution info Use at an assay dependent concentration. | Notes - |
PredictedPredicted
| Species | Dilution info | Notes |
|---|---|---|
Species Chicken, Hamster, Cow, Orangutan | Dilution info - | Notes - |
IHC-P
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Mouse | Dilution info 1/100 | Notes - |
Species Rat | Dilution info 1/100 | Notes - |
Species Human | Dilution info 1/100 | Notes - |
ExpectedExpected
| Species | Dilution info | Notes |
|---|---|---|
Species Pig | Dilution info Use at an assay dependent concentration. | Notes - |
PredictedPredicted
| Species | Dilution info | Notes |
|---|---|---|
Species Chicken, Hamster, Cow, Orangutan | Dilution info - | Notes - |
ICC/IF
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info 5.00000-10.00000 µg/mL | Notes - |
ExpectedExpected
| Species | Dilution info | Notes |
|---|---|---|
Species Mouse, Pig, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
PredictedPredicted
| Species | Dilution info | Notes |
|---|---|---|
Species Chicken, Hamster, Cow, Orangutan | Dilution info - | Notes - |
Associated Products
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Target data
Function
Chaperonin implicated in mitochondrial protein import and macromolecular assembly. Together with Hsp10, facilitates 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 (PubMed:11422376, PubMed:1346131). The functional units of these chaperonins consist of heptameric rings of the large subunit Hsp60, which function as a back-to-back double ring. In a cyclic reaction, Hsp60 ring complexes bind one unfolded substrate protein per ring, followed by the binding of ATP and association with 2 heptameric rings of the co-chaperonin Hsp10. This leads to sequestration of the substrate protein in the inner cavity of Hsp60 where, for a certain period of time, it can fold undisturbed by other cell components. Synchronous hydrolysis of ATP in all Hsp60 subunits results in the dissociation of the chaperonin rings and the release of ADP and the folded substrate protein (Probable).
Alternative names
HSP60, HSPD1, 60 kDa chaperonin, Chaperonin 60, Heat shock protein 60, Heat shock protein family D member 1, HuCHA60, Mitochondrial matrix protein P1, P60 lymphocyte protein, CPN60, HSP-60, Hsp60
Recommended products
Anti-Hsp60 antibody (ab46798) is a rabbit polyclonal antibody that is used to detect Hsp60 in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IP, IHC-P, ICC/IF. Suitable for Human, Mouse, Pig, Rat samples.
- Over 130 publications
- Trusted since 2007
Key facts
- Isotype
- IgG
- Host species
- Rabbit
- Storage buffer
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA- Form
- Liquid
- Clonality
- Polyclonal
- Immunogen
- The exact immunogen used to generate this antibody is proprietary information.
- Purification technique
- Affinity purification Immunogen
- 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.
- Concentration
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage duration
- 1-2 weeks
- Appropriate short-term storage conditions
- +4°C
- Appropriate long-term storage conditions
- -20°C
- Aliquoting information
- Upon delivery aliquot
- Storage information
- Avoid freeze / thaw cycle
Notes
Anti-Hsp60 antibody (ab46798) is a rabbit polyclonal antibody and is validated for use in Western Blot (WB), Flow Cytometry (Intra), Flow Cytometry (Flow Cyt), Immunoprecipitation (IP), Immunohistochemistry (IHC-P), Immunocytochemistry/immunofluorescence (ICC/IF) in Human, Mouse, Pig, Rat samples.
What is the molecular weight of Hsp60?
Anti-Hsp60 (ab46798) specifically detects a band for Hsp60 (UniProt: P10809) at a molecular weight of 60kDa.
Trusted by the scientific community
Anti-Hsp60 (ab46798) was first used in a scientific publication in 2007 and has been cited over 130 times in peer-reviewed journals.
Reviewed by scientists
Anti-Hsp60 (ab46798) has over 10 independent reviews from customers.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Hsp60 also known as HSPD1 and heat shock protein 60 is a significant chaperonin with a molecular weight of approximately 60 kDa. It resides predominantly in the mitochondria and functions to assist in the proper folding of proteins preventing their aggregation. Hsp60 is expressed mainly in cells with high metabolic activity. Its presence as a mitochondrial marker highlights its essential role in maintaining cellular homeostasis and function.
- Biological function summary
The protein ensures mitochondrial protein stability by facilitating the refolding of misfolded proteins and cooperating with other chaperonins like Hsp10. Hsp60 participates in forming a complex with these proteins to create a conducive environment for protein folding. It plays a part in regulating mitochondrial homeostasis impacting cell survival and apoptosis processes.
- Pathways
Hsp60 links to the ATP synthesis and apoptosis pathways showcasing its importance as a mitochondrial marker. It interacts with proteins like caspase-3 to modulate cell death mechanisms highlighting its influence beyond simple protein folding. In the ATP synthesis pathway it contributes indirectly to energy production by maintaining mitochondrial function.
- Associated diseases and disorders
Hsp60 shows a connection to neurodegenerative diseases and cancer. Altered Hsp60 levels correlate with increased apoptosis in neurodegenerative conditions like Alzheimer's disease. Additionally in cancer interactions with proteins such as AKT suggest its potential role in cell proliferation and survival. Understanding its role could aid in the development of therapeutic interventions targeting mitochondrial dysfunction.
Product promise
Tested
We have tested this species and application combination and it works. It is covered by our product promise.
Expected
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
Predicted
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
Not recommended
We do not recommend this combination. It is not covered by our product promise.
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
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16 product images
Western blot - Anti-Hsp60 antibody - Mitochondrial Marker (ab46798)
Hsp60 Western blot staining using rabbit Anti-Hsp60 antibody
Blocking Buffer: 5% NFDM/TBST
Dilution Buffer: 5% NFDM/TBST
All lanes: Western blot - Anti-Hsp60 antibody - Mitochondrial Marker (ab46798) at 1/20000 dilution
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
SecondaryAll lanes: HRP conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 61 kDa
Observed band size: 60 kDa
This image is courtesy of a customer review submitted by Nicole Schechter Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp60 antibody - Mitochondrial Marker (ab46798)
Hsp60 Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining using rabbit Anti-Hsp60 antibody
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.
Flow Cytometry (Intracellular) - Anti-Hsp60 antibody - Mitochondrial Marker (ab46798)
Hsp60 Flow Cytometry (Intracellular) staining using rabbit Anti-Hsp60 antibody
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).
Immunoprecipitation - Anti-Hsp60 antibody - Mitochondrial Marker (ab46798)
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.
All lanes: Immunoprecipitation - Anti-Hsp60 antibody - Mitochondrial Marker (ab46798)
Predicted band size: 61 kDa
Observed band size: 60 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp60 antibody - Mitochondrial Marker (ab46798)
Hsp60 Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining using rabbit Anti-Hsp60 antibody
Immunohistochemical staining of paraffin embedded human lung adenocarcinoma with purified ab46798 at a working dilution of 1 in 100. The secondary antibody used is Goat Anti-Rabbit IgG H&L (HRP) 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.
Western blot - Anti-Hsp60 antibody - Mitochondrial Marker (ab46798)
Hsp60 Western blot staining of T47-D cell lysate using rabbit Anti-Hsp60 antibody
Blocking Buffer: 5% NFDM/TBST
Dilution Buffer: 5% NFDM/TBST
All lanes: Western blot - Anti-Hsp60 antibody - Mitochondrial Marker (ab46798) at 1/20000 dilution
All lanes: T47-D cell lysate at 10 µg
SecondaryAll lanes: HRP conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 61 kDa
Observed band size: 60 kDa
Western blot - Anti-Hsp60 antibody - Mitochondrial Marker (ab46798)
Hsp60 Western blot staining using rabbit Anti-Hsp60 antibody
Blocking Buffer: 5% NFDM/TBST
Dilution Buffer: 5% NFDM/TBST
All lanes: Western blot - Anti-Hsp60 antibody - Mitochondrial Marker (ab46798) at 1/50000 dilution
Lane 1: MCF7 cell lysate at 20 µg
Lane 2: SW480 cell lysate at 20 µg
SecondaryAll lanes: HRP conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 61 kDa
Observed band size: 60 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp60 antibody - Mitochondrial Marker (ab46798)
Hsp60 Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining using rabbit Anti-Hsp60 antibody
Immunohistochemical staining of paraffin embedded rat kidney with purified ab46798 at a working dilution of 1 in 100. The secondary antibody used is Goat Anti-Rabbit IgG H&L (HRP) 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp60 antibody - Mitochondrial Marker (ab46798)
Hsp60 Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining using rabbit Anti-Hsp60 antibody
Immunohistochemical staining of paraffin embedded mouse cardiac muscle with purified ab46798 at a working dilution of 1 in 100. The secondary antibody used is Goat Anti-Rabbit IgG H&L (HRP) 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp60 antibody - Mitochondrial Marker (ab46798)
Hsp60 Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining using rabbit Anti-Hsp60 antibody
Immunohistochemical staining of paraffin embedded human cervical carcinoma with purified ab46798 at a working dilution of 1 in 100. The secondary antibody used is Goat Anti-Rabbit IgG H&L (HRP) 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp60 antibody - Mitochondrial Marker (ab46798)
Hsp60 Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining using rabbit Anti-Hsp60 antibody
Immunohistochemical analysis of paraffin-embedded human breast adenocarcinoma using unpurified ab46798 at 1/250 dilution.
Western blot - Anti-Hsp60 antibody - Mitochondrial Marker (ab46798)
Hsp60 Western blot staining of Pig heart tissue lysate using rabbit Anti-Hsp60 antibody
Blocking Buffer: 5% NFDM/TBST
Dilution Buffer: 5% NFDM/TBST
All lanes: Western blot - Anti-Hsp60 antibody - Mitochondrial Marker (ab46798) at 1/50000 dilution
All lanes: Pig heart tissue lysate at 20 µg
SecondaryAll lanes: HRP conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 61 kDa
Observed band size: 60 kDa
Western blot - Anti-Hsp60 antibody - Mitochondrial Marker (ab46798)
Hsp60 Western blot staining of T-47D whole cell lysate using rabbit Anti-Hsp60 antibody
Secondary antibody - anti-rabbit HRP (Goat Anti-Rabbit IgG H&L (HRP) ab6721)
All lanes: Western blot - Anti-Hsp60 antibody - Mitochondrial Marker (ab46798) at 1/50000 dilution
All lanes: Western blot - T-47D whole cell lysate (T-47D whole cell lysate ab14899) at 10 µg
SecondaryAll lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab6721) at 1/2000 dilution
Predicted band size: 61 kDa
Observed band size: 60 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp60 antibody - Mitochondrial Marker (ab46798)
Hsp60 Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining of Human liver using rabbit Anti-Hsp60 antibody
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
This image is courtesy of a customer review submitted by Suleyman BozkurtImmunocytochemistry - Anti-Hsp60 antibody - Mitochondrial Marker (ab46798)
Hsp60 Immunocytochemistry staining of HeLa FlipIn Cell line using rabbit Anti-Hsp60 antibody
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.
Immunocytochemistry/ Immunofluorescence - Anti-Hsp60 antibody - Mitochondrial Marker (ab46798)
Hsp60 Immunocytochemistry/ Immunofluorescence staining of HeLa cells using rabbit Anti-Hsp60 antibody
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 Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed 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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