Antibodies, Proteins, Kits and Reagents for Life Science

291 484 ₸ Product size

100 µl 291 484 ₸ 10 µl 130 665 ₸

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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, Flow Cyt, IP
Reacts with: Mouse, Rat, Human

Product name

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

Rabbit monoclonal [EPR18245-93] to Hsp60
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Mouse
ICC/IF	Mouse Human
IHC-P	Mouse Rat Human
IP	Mouse
WB	Mouse Rat Human

See all applications and species data

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: 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

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

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 Fluor^® 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)

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