Overview

  • Product name
    Anti-Hsp60 antibody [Mab11-13]
    See all Hsp60 primary antibodies
  • Description
    Mouse monoclonal [Mab11-13] to Hsp60
  • Host species
    Mouse
  • Tested applications
    Suitable for: WB, IP, Flow Cyt, Electron Microscopy, ICC/IF, IHC-Fr, ELISAmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Rabbit, Guinea pig, Hamster, Cow, Dog, Human, Pig, Drosophila melanogaster, Monkey, Snake, Dolphin, Rainbow trout
    Does not react with: Saccharomyces cerevisiae, Escherichia coli
  • Immunogen

    Recombinant fragment, amino acids 31-547 (Human).

  • Epitope
    This antibody recognizes a surface epitope of Hsp60 in the region of amino acids 288-366.
  • Positive control
    • HeLa Heat Shocked Cell Lysate or Recombinant Human Hsp60 Protein IF/ICC: HeLa cell line.
  • General notes

    Abcam is committed to meeting high standards of ethical manufacturing and as such, we will be discontinuing this product, which has been generated by the ascites method, within the next year. We are sorry for any inconvenience this may cause. If you would like help finding an alternative product, please do not hesitate to contact our scientific support team.

Properties

Applications

Our Abpromise guarantee covers the use of ab13532 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 1/10000. Detects a band of approximately 60 kDa (predicted molecular weight: 68.8 kDa).
IP Use at an assay dependent concentration.
Flow Cyt Use 0.5µg for 106 cells.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

 

Electron Microscopy Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.

Target

  • 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
  • Database links
  • 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

Images

  • ICC/IF image of ab13532 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab13532, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse IgG(H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Overlay histogram showing HeLa cells stained with ab13532 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab13532, 0.5µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

References

This product has been referenced in:
  • Vais H  et al. EMRE Is a Matrix Ca(2+) Sensor that Governs Gatekeeping of the Mitochondrial Ca(2+) Uniporter. Cell Rep 14:403-410 (2016). Read more (PubMed: 26774479) »
  • Zhu Z  et al. Dynamics of the Transcriptome during Human Spermatogenesis: Predicting the Potential Key Genes Regulating Male Gametes Generation. Sci Rep 6:19069 (2016). Read more (PubMed: 26753906) »
See all 10 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Answer

Thank you for your reply. I have requested that a vial of ab13532 be added free of charge to your next order. I apologize for any inconveniences caused to the customer. Please do not hesitate to contact me if you have any additional questions.

Read More

Question

Dear Tech Support Team, Please see below the details of customer's complaint. Attached are the images. Thanks in advance for your assistance and reply. General Information: Antibody storage conditions (temperature/reconstitution etc) Aliquot store at -20deg C. Description of the problem: we used this antibody for the last 5 years and for the first time this specific lot didn't work as expected, when we used this antibody in our regular protocol for Immunocytochemistry the staining was not good (as can be seen in the added pic). We used this antibody (other LOTs) and it worked great! So we kept the same amount of antibody as we used before. Sample (Species/Tissue/Cell Type/Cell Line etc.) fibrosarcoma cell line and NIH-3T3 cell line Fixation of sample (Ethanol/Methanol/Acetone/Paraformaldehyde/Other/Duration etc.) paraformaldehyde Permeabilization step - Triton Blocking conditions (Buffer/time period, Blocking agent etc.) 1% BSA Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step) Dylight 549 donkey anti mouse Detection method confocal microscopy Positive and negative controls used (please specify) The important control for this complaint is the pic of the same antibody in the same protocol just old lot, and the staining with this antibody was good. Optimization attempts (problem solving): How many times have you tried the ICC? Just once with this LOT and a lot more with other lots of the same antibody Have you run a No Primary control? no Do you obtain the same results every time? yes What steps have you altered? Non

Read More
Answer

Thank you for contacting Abcam regarding ab13532. I am sorry that the lot the customer has just received of this antibody is not working as well as previous lots. I would be happy to offer a replacement for the customer. Can you please provide the order number? I look forward to your reply so that I may assist you further. Please do not hesitate to contact me if you have any additional questions.

Read More

Answer

Thank you for your enquiry and for completing our technical questionnaire. I am sorry to hear that you have been having difficulties with this antibody. I have read your questionnaire and I have a few comments. When you state that you used a positive control antibodies to detect mitochondrial proteins can you please tell me whether you were successful or not. For the technique of immunocytochemistry the permeabilization is crucial. I would therefore like to recommend that you perform a 30-min incubation in PBS containing 0.5% Triton X-100 as detailed in the methods section of Satoh et al., (1998). In the paper they have applied a hsp60 antibody using fibroblasts and whilst they have not used an Abcam antibody they have presented the antigen successfully to the antibody using these permeabilization conditions. Satoh J, Yukitake M, Kurohara K, Nishida N, Katamine S, Miyamoto T, Kuroda Y. (1998) Cultured skin fibroblasts isolated from mice devoid of the prion protein gene Express major heat shock proteins in response to heat stress. Exp Neurol. 151(1):105-15. PMID: 9582258 I would like to recommend that you supplement this permeabilization step into our ICC protocol which differs slightly from the protocol that you have detailed by fixation using paraformaldehyde rather than formaldehyde and washes using PBST. It can be accessed at the following link: https://www.abcam.com/assets/pdf/protocols/Immunocytochemistry%20(ICC)%20protocol.pdf I would also like to recommend that you perform a series of dilutions to determine whether you can obtain any staining. Unfortunately we do not have details of the optimal dilution and this should be determined empirically and future experiments adjusted accordingly. I will contact the source of this antibody to determine whether it has been quantitated. However, given that it is a monoclonal antibody it is predicted to be between 0.5 and 1mgml-1. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

Read More

Question
Answer

This antibody goes only by volume even though it is purified protein. The amount is proprietary information.

Read More

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Sign up