Anti-Hsp60 antibody [4B9/89]

• 产品名称

  Anti-Hsp60抗体[4B9/89]
  参阅全部 Hsp60 一抗

• 描述

  小鼠单克隆抗体[4B9/89] to Hsp60

• 宿主

  Mouse

• 经测试应用

  适用于: Flow Cyt, IHC-P, ICC/IF, IP, WBmore details

• 种属反应性

  与反应: Mouse, Human
  

• 免疫原

  Full length protein corresponding to Human Hsp60. Human placental Hsp60.

• 表位

  Epitope mapping studies using human Hsp 60 deletion mutants suggest that this antibody binds either between amino acids 335-366 or 484-547.

• 常规说明

  
  

  The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

  If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

• 形式

  Liquid

• 存放说明

  Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.

• 存储溶液

  Constituent: 100% PBS

• 浓度

• 100 µg 浓度为 1 mg/ml

• 纯度

  Protein A purified

• 克隆

  单克隆

• 克隆编号

  4B9/89

• 同种型

  IgG2a

The Abpromise guarantee

Abpromise™承诺保证使用ab5478于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

• 数据库链接

• Entrez Gene: 3329 Human

• Entrez Gene: 15510 Mouse

• Omim: 118190 Human

• SwissProt: P10809 Human

• SwissProt: P63038 Mouse

• Unigene: 595053 Human

• Unigene: 727543 Human

• Unigene: 1777 Mouse

• 别名

  hide

• 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

• 60 kDa chaperonin antibody

• 60 kDa heat shock protein, mitochondrial antibody

• CH60_HUMAN antibody

• 

  Western blot - Anti-Hsp60 antibody [4B9/89] (ab5478)

  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
  All lanes : HRP-conjugated goat anti-mouse IgG at 1/20000 dilution
  

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Hsp60 antibody [4B9/89] (ab5478)

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

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

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

  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.

• 

  Flow Cytometry - Anti-Hsp60 antibody [4B9/89] (ab5478)

  Flow cytometry analysis of HSP60 was done on HeLa cells. Cells were fixed, permeabilized and stained with a HSP60 mouse monoclonal antibody (orange histogram) or a mouse IgG2a isotype control (black histogram) at a dilution of 10 µg/mL. After incubation for 1 hour on ice, the cells were labeled with a Goat anti-Mouse IgG Secondary Antibody, DyLight 650 conjugate at 1/50 dilution for 1 hour on ice. A representative 10,000 cells were acquired and analyzed for each sample.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Hsp60 antibody [4B9/89] (ab5478)

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

  Immunoprecipitation of Hsp60 was performed on HeLa cells. Antigen:antibody complexes were formed by incubating 500µg whole cell lysate with 2µg of HSP60 monoclonal antibody (ab5478) overnight on a rocking platform at 4°C. The immune complexes were captured on 50µl 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.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp60 antibody [4B9/89] (ab5478)

  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.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Hsp60 antibody [4B9/89] (ab5478)

  Immunolocalization of Hsp 60 in human endothelial cells using ab5478.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp60 antibody [4B9/89] (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 (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp60 antibody [4B9/89] (ab5478)

  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.