製品の概要

  • 製品名Anti-Hsp60 antibody [LK-1]
    Hsp60 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [LK-1] to Hsp60
  • アプリケーション適用あり: WB, IP, ELISA, Flow Cyt, IHC-Fr, IHC-P, ICC/IFmore details
  • 種交差性
    交差種: Mouse, Rat, Sheep, Rabbit, Chicken, Guinea pig, Hamster, Cow, Dog, Human, Pig, Xenopus laevis, Drosophila melanogaster, Monkey
  • 免疫原

    Human Hsp60 produced through recombinant DNA methods in E.coli

  • ポジティブ・コントロール
    • HeLa cell lysate

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab59457 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
WB Use a concentration of 0.05 µg/ml. Predicted molecular weight: 60 kDa.
IP Use a concentration of 5 µg/ml.
ELISA Use at an assay dependent concentration.
Flow Cyt Use 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
IHC-Fr Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.
ICC/IF 1/100.

ターゲット情報

  • 機能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.
  • 関連疾患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.
  • 配列類似性Belongs to the chaperonin (HSP60) family.
  • 細胞内局在Mitochondrion matrix.
  • Information by UniProt
  • 参照データベース
  • 別名
    • 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

Anti-Hsp60 antibody [LK-1] 画像

  • ICC/IF image of ab59457 stained HepG2 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 (ab59457, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 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.

  • Ab59457 staining human normal skin tissue. Staining is localised to mitochondria.
    Left panel: with primary antibody at 2 ug/ml. Right panel: isotype control.
    Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 antigen retrieval buffer EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
  • All lanes : Anti-Hsp60 antibody [LK-1] (ab59457) at 0.05 µg/ml

    Lane 1 : Rat Brain tissue lysates
    Lane 2 : Rat Heart tissue lysates
    Lane 3 : Rat Kidney tissue lysates
    Lane 4 : Rat Liver tissue lysates
    Lane 5 : Rat Lung tissue lysates
    Lane 6 : Rat Pancreas tissue lysates
    Lane 7 : Rat skeletal muscle tissue lysate
    Lane 8 : Rat Spleen tissue lysate
    Lane 9 : Rat Testes tissue lysate
    Lane 10 : Rat Thymus tisuue lysate
    Lane 11 : Cell lysates prepared from rat heart H9C2 cells
    Lane 12 : Cell lysates prepared from mouse NIH3T3 cells
    Lane 13 : Cell lysates prepared from mouse Pam212 cells

    Lysates/proteins at 10 µg per lane.

    Secondary
    HRP-conjugated goat polyclonal to mouse IgG1 at 1/10 dilution

    Predicted band size : 60 kDa
  • Overlay histogram showing HeLa cells stained with ab59457 (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 (ab59457, 1µg/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 IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • Hsp60 was immunoprecipitated using 0.5mg Rat Brain tissue lysate, 5µg of Mouse monoclonal to Hsp60 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Rat Brain tissue lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab59457.
    Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/20,000 dilution.
    Band: 60kDa; Hsp60: non specific bands - 50kDa: We are unsure as to the identity of this extra band.

Anti-Hsp60 antibody [LK-1] (ab59457) 使用論文

This product has been referenced in:
  • Rupprecht A  et al. Uncoupling protein 2 and 4 expression pattern during stem cell differentiation provides new insight into their putative function. PLoS One 9:e88474 (2014). WB ; Mouse . Read more (PubMed: 24523901) »
  • Mizumura K  et al. Mitophagy-dependent necroptosis contributes to the pathogenesis of COPD. J Clin Invest 124:3987-4003 (2014). Read more (PubMed: 25083992) »

See all 4 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Colon malignant)
Specification Colon malignant
Fixative Paraformaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Novocastra pH 6 retrieval buffer
Permeabilization No
Blocking step H2O2 as blocking agent for 10 minute(s) · Concentration: 3% · Temperature: 22°C
Username

Dr. Aamir Ahmed

Verified customer

投稿 Jan 25 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (SHSY5Y)
Loading amount 50 µg
Specification SHSY5Y
Treatment LPS
Gel Running Conditions Reduced Denaturing (12%)
Blocking step Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 Jan 20 2011

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