HSP60 Antibody - BSA Free

Novus Biologicals | Catalog # NBP1-77397

Novus Biologicals
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Key Product Details

Species Reactivity

Validated:

Human, Mouse

Cited:

Human, Mouse

Predicted:

Bovine (99%), Chicken (100%), Hamster (98%), Orangutan (100%), Rat (100%). Backed by our 100% Guarantee.

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Immunoblotting, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Simple Western, Knockdown Validated

Cited:

Western Blot, Immunoblotting, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Knockdown Validated

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all HSP60 Primary Antibodies »

Product Specifications

Immunogen

A synthetic peptide made to an internal region of the human Hsp60 protein (within residues 70-150). [Swiss-Prot P10809]

Localization

Mitochondrion matrix

Marker

Mitochondria Marker

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for HSP60 Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: HSP60 Antibody - BSA Free [NBP1-77397]

Immunocytochemistry/ Immunofluorescence: HSP60 Antibody - BSA Free [NBP1-77397]

Immunocytochemistry/Immunofluorescence: HSP60 Antibody [NBP1-77397] - HeLa cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with HSP60 Antibody (NBP1-77397) at 1 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.
Western Blot: HSP60 AntibodyBSA Free [NBP1-77397]

Western Blot: HSP60 AntibodyBSA Free [NBP1-77397]

Western Blot: HSP60 Antibody [NBP1-77397] - Analysis of extracts from HeLa and COS cells using NBP1-77397 Hsp60 antibody at 1:1000
Immunocytochemistry/ Immunofluorescence: HSP60 Antibody - BSA Free [NBP1-77397]

Immunocytochemistry/ Immunofluorescence: HSP60 Antibody - BSA Free [NBP1-77397]

Immunocytochemistry/Immunofluorescence: HSP60 Antibody - BSA Free [NBP1-77397] - Rat FR cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with HSP60 Antibody (NBP1-77397) at 1ug/ml overnight at 4C and detected with an anti-rabbit DyLight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.
Immunohistochemistry: HSP60 Antibody - BSA Free [NBP1-77397]

Immunohistochemistry: HSP60 Antibody - BSA Free [NBP1-77397]

Immunohistochemistry: HSP60 Antibody [NBP1-77397] - Staining of HSP60 in human kidney carcinoma using DAB with hematoxylin counterstain.
Flow Cytometry: HSP60 Antibody - BSA Free [NBP1-77397]

Flow Cytometry: HSP60 Antibody - BSA Free [NBP1-77397]

Flow Cytometry: HSP60 Antibody [NBP1-77397] - Analysis of HSP60 in RM1 cells (murine prostate cancer cell line) using anti-HSP60 antibody. The primary antibody was used at a dilution of 1:100. Image from verified customer review.
Western Blot: HSP60 AntibodyBSA Free [NBP1-77397]

Western Blot: HSP60 AntibodyBSA Free [NBP1-77397]

Western Blot: HSP60 Antibody [NBP1-77397] - Analysis of HSP60 in: 1) HeLa, 2) HepG2, 3) NIH/3T3, 4) Jurkat, 5) CHO, 6) A431, 7) PC12 and 8) COS7
Immunocytochemistry/ Immunofluorescence: HSP60 Antibody - BSA Free [NBP1-77397]

Immunocytochemistry/ Immunofluorescence: HSP60 Antibody - BSA Free [NBP1-77397]

Immunocytochemistry/Immunofluorescence: HSP60 Antibody [NBP1-77397] - HSP60 antibody was tested in HeLa cells with FITC (green). Nuclei and actin were counterstained with DAPI (blue) and Phalloidin (red).
Immunocytochemistry/ Immunofluorescence: HSP60 Antibody - BSA Free [NBP1-77397]

Immunocytochemistry/ Immunofluorescence: HSP60 Antibody - BSA Free [NBP1-77397]

Immunocytochemistry/Immunofluorescence: HSP60 Antibody [NBP1-77397] - Confocal immunofluorescence analysis of HeLa cells using Hsp60 antibody at 1:50 (green). Nuclei were counterstained using DAPI (blue).
Immunocytochemistry/ Immunofluorescence: HSP60 Antibody - BSA Free [NBP1-77397]

Immunocytochemistry/ Immunofluorescence: HSP60 Antibody - BSA Free [NBP1-77397]

Immunocytochemistry/Immunofluorescence: HSP60 Antibody [NBP1-77397] - Confocal immunofluorescent analysis of HeLa cells using Hsp60 antibody (NBP1-77397, 1:100). An Alexa Fluor 488-conjugated Goat to rabbit IgG was used as secondary antibody (green, A). Actin filaments were labeled with Alexa Fluor 568 phalloidin (red, B). DAPI was used to stain the cell nuclei (blue, C).
Immunocytochemistry/ Immunofluorescence: HSP60 Antibody - BSA Free [NBP1-77397]

Immunocytochemistry/ Immunofluorescence: HSP60 Antibody - BSA Free [NBP1-77397]

Immunocytochemistry/Immunofluorescence: HSP60 Antibody [NBP1-77397] - HeLa cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with HSP60 Antibody conjugated to Alexa Fluor 488 (NBP1-77397AF488) at 5 ug/ml for 1 hour at room temperature. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.
Immunocytochemistry/ Immunofluorescence: HSP60 Antibody - BSA Free [NBP1-77397]

Immunocytochemistry/ Immunofluorescence: HSP60 Antibody - BSA Free [NBP1-77397]

Immunocytochemistry/Immunofluorescence: HSP60 Antibody - BSA Free [NBP1-77397] - Mouse MS1 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with HSP60 Antibody (NBP1-77397) at 2ug/ml overnight at 4C and detected with an anti-rabbit DyLight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.
Immunocytochemistry/ Immunofluorescence: HSP60 Antibody - BSA Free [NBP1-77397]

Immunocytochemistry/ Immunofluorescence: HSP60 Antibody - BSA Free [NBP1-77397]

Immunocytochemistry/Immunofluorescence: HSP60 Antibody - BSA Free [NBP1-77397] - HepG2 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with HSP60 Antibody (NBP1-77397) at 1ug/ml overnight at 4C and detected with an anti-rabbit DyLight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.
Simple Western: HSP60 AntibodyBSA Free [NBP1-77397]

Simple Western: HSP60 AntibodyBSA Free [NBP1-77397]

Simple Western: HSP60 Antibody [NBP1-77397] - Lane view shows a specific band for Hsp60 in 0.05 mg/ml of HepG2 lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Applications for HSP60 Antibody - BSA Free

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:50-1:1000

Immunohistochemistry

1:100

Immunohistochemistry-Paraffin

1:100

Knockdown Validated

reported in scientific literature (Shi et al)

Simple Western

1:5000

Western Blot

0.5 ug/ml
Application Notes
In Western Blot, a band is seen ~61 kDa representing Hsp60. Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (pH 6.0) is recommended. In Simple Western only 10-15 uL of the recommended dilution is used per data point.
See Simple Western Antibody Database for Simple Western validation: Tested in HepG2 lysate 0.05 mg/mL, separated by Size, antibody dilution of 1:5000, apparent MW was 62 kDa. Separated by Size-Wes, Sally Sue/Peggy Sue.

Reviewed Applications

Read 4 reviews rated 5 using NBP1-77397 in the following applications:

Flow Cytometry Panel Builder

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Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS and 30% Glycerol

Format

BSA Free

Preservative

0.05% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: HSP60

Heat shock protein 60 activates T cells and ultimately autoimmune actions and diseases. In gastric cancer, HSP60 is upregulated in malignant ascites that have been treated, and induces a tumor-specific cytotoxic T lymphocyte response. Enhancing Hsp60 expression is beneficial for protecting bone tissue against osteoblast apoptosis. Hsp60 is highly influenced by dietary intake of antioxidants, and is a prime target for many natural therapies against gut inflammation, preeclampsia, and some low level bacterial infections. Ischemia and reperfusion, as well as hypoxic environments can cause temporary elevations in Hsp60 expression levels.

Long Name

Heat Shock Protein 60

Alternate Names

cpn60, GROEL, HSPD1, SPG13

Entrez Gene IDs

3329 (Human); 15510 (Mouse)

Gene Symbol

HSPD1

UniProt

P63038 (Mouse), P10809 (Mouse)

Additional HSP60 Products

Product Documents for HSP60 Antibody - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Download SDS

Product Specific Notices for HSP60 Antibody - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Related Research Areas

Heat Shock Proteins (HSPs)

Citations for HSP60 Antibody - BSA Free

Customer Reviews for HSP60 Antibody - BSA Free (4)

5 out of 5
4 Customer Ratings
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4 Stars
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Customer Images

Showing  1 - 4 of 4 reviews Showing All
Filter By:
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  • Name: Anonymous
    Application: Flow Cytometry
    Sample Tested: Murine prostate cancer cell line RM1
    Species: Mouse
    Verified Customer | Posted 12/30/2014
    Anti-Hsp60 antibody (NBP1 77397SS) tested on murine prostate cancer cells (RM1).
    HSP60 Antibody - BSA Free NBP1-77397
  • Name: Bryan Tinsley
    Application: Immunocytochemistry
    Sample Tested:
    Species: Human
    Verified Customer | Posted 06/05/2014
    Confocal immunofluorescent analysis of HeLa cells using Hsp60 antibody (NBP1-77397, 1:100).
    HSP60 Antibody - BSA Free NBP1-77397
  • Name: Bryan Tinsley
    Application: Western Blot
    Sample Tested:
    Species: Human
    Verified Customer | Posted 06/05/2014
    HeLa and COS cells using Hsp60 antibody (NBP1-77397, 1:2000).
    HSP60 Antibody - BSA Free NBP1-77397
  • Name: Bryan Tinsley
    Application: Immunocytochemistry
    Sample Tested:
    Species: Human
    Verified Customer | Posted 05/21/2014
    Confocal immunofluorescence analysis of HeLa cells using Hsp60 antibody at 1:50 (green). Nuclei were counterstained using DAPI.

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Showing  1 - 4 of 4 reviews Showing All

Protocols

View specific protocols for HSP60 Antibody - BSA Free (NBP1-77397):


Immunocytochemistry Protocol

Culture cells to appropriate density in 35 mm culture dishes or 6-well plates.

1. Remove culture medium and add 10% formalin to the dish. Fix at room temperature for 30 minutes.
2. Remove the formalin and add ice cold methanol. Incubate for 5-10 minutes.
3. Remove methanol and add washing solution (i.e. PBS). Be sure to not let the specimen dry out. Wash three times for 10 minutes.
4. To block nonspecific antibody binding incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
5. Add primary antibody at appropriate dilution and incubate at room temperature from 2 hours to overnight at room temperature.
6. Remove primary antibody and replace with washing solution. Wash three times for 10 minutes.
7. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
8. Remove antibody and replace with wash solution, then wash for 10 minutes. Add Hoechst 33258 to wash solution at 1:25,0000 and incubate for 10 minutes. Wash a third time for 10 minutes.
9. Cells can be viewed directly after washing. The plates can also be stored in PBS containing Azide covered in Parafilm (TM). Cells can also be cover-slipped using Fluoromount, with appropriate sealing.

*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.


Immunohistochemistry-Paraffin Embedded Sections

Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes.

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in wash buffer for 5 minutes.
3. Block each section with 100-400 ul blocking solution for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul biotinylated diluted secondary antibody. Incubate 30 minutes at room temperature.
7. Remove secondary antibody solution and wash sections three times with wash buffer for 5 minutes each.
8. Add 100-400 ul Streptavidin-HRP reagent to each section and incubate for 30 minutes at room temperature.
9. Wash sections three times in wash buffer for 5 minutes each.
10. Add 100-400 ul DAB substrate to each section and monitor staining closely.
11. As soon as the sections develop, immerse slides in deionized water.
12. Counterstain sections in hematoxylin.
13. Wash sections in deionized water two times for 5 minutes each.
14. Dehydrate sections.
15. Mount coverslips.


Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 40 ug of total protein per lane.
2. Transfer proteins to membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain according to standard Ponceau S procedure (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot.
5. Block the membrane using standard blocking buffer for at least 1 hour.
6. Wash the membrane in wash buffer three times for 10 minutes each.
7. Dilute primary antibody in blocking buffer and incubate 1 hour at room temperature.
8. Wash the membrane in wash buffer three times for 10 minutes each.
9. Apply the diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.
10. Wash the blot in wash buffer three times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturers instructions.

*Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2%.

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FAQs

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