製品の概要

  • 製品名Anti-Hsp60 antibody [4B9/89]
    Hsp60 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [4B9/89] to Hsp60
  • アプリケーション適用あり: Inhibition Assay, ICC/IF, IP, ELISA, ICC, WBmore details
  • 種交差性
    交差種: Mouse, Rat, Hamster, Human, Non Human Primates
  • 免疫原

    Other Immunogen Type corresponding to Human Hsp60. Human placental Hsp60.

  • エピトープEpitope mapping studies using human Hsp 60 deletion mutants suggest that this antibody binds either between amino acids 335-366 or 484-547.
  • ポジティブ・コントロール
    • WB: human blood. ICC: B-SC-1 cells.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab5478 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
Inhibition Assay Use at an assay dependent concentration.
ICC/IF 1/50.
IP Use a concentration of 2 µg/ml.
ELISA Use at an assay dependent concentration.
ICC Use a concentration of 10 µg/ml.

Immunocytochemical staining of Hsp 60 in B-SC-1 cells with ab5478 yields a pattern consistent with the mitochondrial localization of Hsp 60.

WB 1/100 - 1/1000. Detects a band of approximately 60 kDa.

Detects a band of approximately 60 kDa representing Hsp 60 from human blood samples.

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 [4B9/89] 画像

  • Immunocytochemistry/Immunofluorescence analysis of Hsp60 in A2058 Cells. Hsp60 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with ab5478 at a dilution of 1:200 over night at 4 ?C, washed with PBS and incubated with a DyLight-488 conjugated goat anti-mouse IgG secondary antibody. Images were taken at 60X magnification.

  • Immunocytochemistry/Immunofluorescence analysis of Hsp60 in ATDC5 Cells. Hsp60 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were incubated without (control) or with ab5478 at a dilution of 1:100 over night at 4°C, washed with PBS and incubated with a DyLight-488 conjugated goat anti-mouse IgG secondary antibody. Images were taken at 60X magnification.

  • Immunocytochemistry/Immunofluorescence analysis of Hsp60 in Hela Cells. Hsp60 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were incubated without (control) or with ab5478 at a dilution of 1:100 over night at 4°C, washed with PBS and incubated with a DyLight-488 conjugated goat anti-mouse secondary antibody. Images were taken at 60X magnification.

  • Immunocytochemistry/Immunofluorescence analysis of Hsp60 (green) in HeLa cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with (left panel) or without (right panel) ab5478 at a dilution of 1:50 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat-anti-mouse IgG secondary antibody at a dilution of 1:400 for 30 minutes at room temperature. F-Actin (red) was stained with Dylight 554 phalloidin, and nuclei (blue) were stained with Hoechst 33342 dye. Images were taken at 20X magnification.

  • Immunocytochemistry/Immunofluorescence analysis of Hsp60 (green) in HeLa and NIH3T3 cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with ab5478 at a dilution of 1:50 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat-anti-mouse IgG secondary antibody at a dilution of 1:400 for 30 minutes at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye. Images were taken at 20X magnification.

  • Immunoprecipitation of Hsp60 was performed on HeLa cells. Antigen:antibody complexes were formed by incubating 500�g whole cell lysate with 2�g of HSP60 monoclonal antibody (ab5478) overnight on a rocking platform at 4�C. The immune complexes were captured on 50�l Protein Agarose washed extensively and eluted with Buffer. Samples were then resolved on a 4-20% Tris-HCl polyacrylamide gel then transferred to a PVDF membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with a HSP60 monoclonal antibody (ab5478) at a dilution of 1:1000 overnight rotating at 4�C, washed in TBSTand probed with Detection Reagent (HRP) at a dilution of 1:1000 for at least one hour. Chemiluminescent detection was performed.
  • All lanes : Anti-Hsp60 antibody [4B9/89] (ab5478) at 1/1000 dilution

    Lane 1 : 293T cell lysate
    Lane 2 : HeLa cell lysate
    Lane 3 : K562 cell lysate
    Lane 4 : A431 cell lysate
    Lane 5 : HepG2 cell lysate

    Lysates/proteins at 50 µg per lane.

    Secondary
    HRP-conjugated goat anti-mouse IgG at 1/20000 dilution
  • Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized human Kidney tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:100 with a mouse monoclonal antibody recognizing Heat Shock Protein 60 (ab5478) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.
  • Immunolocalization of Hsp 60 in human endothelial cells using ab5478.
  • Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized human Tonsil tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:100 with a mouse monoclonal antibody recognizing Heat Shock Protein 60 (ab5478) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.
  • Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized human Breast carcinoma tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:50 with a mouse monoclonal antibody recognizing Heat Shock Protein 60 (ab5478) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

Anti-Hsp60 antibody [4B9/89] (ab5478) 使用論文

This product has been referenced in:
  • Liang X  et al. Pinch1 is required for normal development of cranial and cardiac neural crest-derived structures. Circ Res 100:527-35 (2007). IHC-P ; Mouse . Read more (PubMed: 17272814) »

See 1 Publication for this product

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