Anti-Hsp60 antibody [2E1/53]

• 产品名称

  Anti-Hsp60抗体[2E1/53]
  参阅全部 Hsp60 一抗

• 描述

  小鼠单克隆抗体[2E1/53] to Hsp60

• 宿主

  Mouse

• 特异性

  ab5479 detects Hsp 60 from Human cells, tissues and recombinant protein preparations. This antibody displays no other protein or species cross-reactivity.

• 经测试应用

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

• 种属反应性

  与反应: Mouse, Rat, Human, African green monkey
  

• 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 between amino acids 211-288.

• 阳性对照

• WB: HeLa ,A431, HEK-293, HepG2, COS-7, NIH/3T3, THP1, K562, A549, HEK-293T, U-2 OS, NRK whole cell lysate. IHC-P: Human kidney and breast carcinoma tissue. ICC: U-251, A431, HeLa, NIH-3T3 cells. IP: HeLa whole cell lysates.

• 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

  Preservative: 0.05% Sodium azide
  Constituents: 0.1% BSA, 99% PBS

• 浓度

• 100 µg 浓度为 1 mg/ml

• 纯度

  Purified IgM

• 克隆

  单克隆

• 克隆编号

  2E1/53

• 同种型

  IgM

The Abpromise guarantee

Abpromise™承诺保证使用ab5479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

• Entrez Gene: 63868 Rat

• Omim: 118190 Human

• SwissProt: P10809 Human

• SwissProt: P63038 Mouse

• SwissProt: P63039 Rat

• Unigene: 595053 Human

• Unigene: 727543 Human

• Unigene: 1777 Mouse

• Unigene: 102058 Rat

别名

• 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

• 

  Western blot - Anti-Hsp60 antibody [2E1/53] (ab5479)

  All lanes : Anti-Hsp60 antibody [2E1/53] (ab5479) at 1/2000 dilution
  
  Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
  Lane 2 : A431 (human epidermoid carcinoma cell line) whole cell lysate
  Lane 3 : HEK-293 (h
  uman epithelial cell line from embryonic kidney) whole cell lysate
  Lane 4 : HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate
  Lane 5 : COS-7 (african green monkey kidney fibroblast-like cell line) whole cell lysate
  Lane 6 : NIH/3T3 (mouse embryonic fibroblast cell line) whole cell lysate
  Lane 7 : THP1 (human monocytic leukemia cell line) whole cell lysate
  Lane 8 : K562 (human chronic myelogenous leukemia lymphoblast cell line ) whole cell lysate
  Lane 9 : A549 (human lung carcinoma cell line) whole cell lysate
  
  Lysates/proteins at 30 µg per lane.
  
  Secondary
  All lanes : Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate at 1/4000 dilution
  

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Hsp60 antibody [2E1/53] (ab5479)

  Immunofluorescence analysis of HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling HSP60 at 1/200 dilution in 0.1% BSA, incubated at 4°C overnight, followed by Goat anti-Mouse IgG (H+L)/IgM (L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate at 1/2000 dilution for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI. F-actin (Panel c: red) was stained with Rhodamine Phalloidin at 1/300 dilution. Panel d represents the merged image showing mitochondrial localization. Panel e represents control cells with no primary antibody to assess background. 

  The cells (70% confluent log phase) were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The images were captured at 60X magnification.

• 

  Western blot - Anti-Hsp60 antibody [2E1/53] (ab5479)

  All lanes : Anti-Hsp60 antibody [2E1/53] (ab5479) at 1 µg/ml (overnight at 4°C)
  
  Lane 1 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate with 5% BSA/TBST for at least 1 hour
  Lane 2 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate with 5% BSA/TBST for at least 1 hour
  Lane 3 : K562 (human chronic myelogenous leukemia lymphoblast cell line ) whole cell lysate with 5% BSA/TBST for at least 1 hour
  Lane 4 : A431 (human epidermoid carcinoma cell line) whole cell lysate with 5% BSA/TBST for at least 1 hour
  Lane 5 : HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate with 5% BSA/TBST for at least 1 hour
  Lane 6 : U-2 OS (human bone osteosarcoma epithelial cell line) whole cell lysate with 5% BSA/TBST for at least 1 hour
  Lane 7 : COS-7 (african green monkey kidney fibroblast-like cell line) whole cell lysate with 5% BSA/TBST for at least 1 hour
  Lane 8 : NIH/3T3 (mouse embryonic fibroblast cell line) whole cell lysate with 5% BSA/TBST for at least 1 hour
  Lane 9 : NRK (rat kidney cell line) whole cell lysate with 5% BSA/TBST for at least 1 hour
  
  Lysates/proteins at 50 µg per lane.
  
  Secondary
  All lanes : Goat anti-mouse IgM secondary antibody at 1/20000 dilution
  
  
  

  Western blot analysis of various cell lines (50µg/lane) labeling Hsp60 with ab5479 at 1µg/ml.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Hsp60 antibody [2E1/53] (ab5479)

  Immunofluorescence analysis of Hsp60 (green) in HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling Hsp60 with ab5479 at 1/200 dilution and incubated overnight in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody for 45 minutes at room temperature in the dark. F-actin (red) was stained with a fluorescent phalloidin and nuclei (blue) were stained with DAPI.

  Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes at room temperature and blocked with 3% BSA-PBS for 30 minutes at room temperature. Images were taken at a 60X magnification.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Hsp60 antibody [2E1/53] (ab5479)

  Immunofluorescence analysis of A431 (human epidermoid carcinoma cell line) cells labeling Hsp60 (green) with ab5479 at 1/100 dilution and incubated overnight in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody for 45 minutes at room temperature in the dark. F-actin (red) was stained with a fluorescent phalloidin and nuclei (blue) were stained with DAPI. 

  Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes at room temperature and blocked with 3% BSA-PBS for 30 minutes at room temperature. Images were taken at a 60X magnification.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Hsp60 antibody [2E1/53] (ab5479)

  Immunofluorescence analysis of NIH/3T3 (mouse embryonic fibroblast cell line) cells labeling Hsp60 (green) with ab5479 at 1/20 dilution and incubated overnight in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody for 45 minutes at room temperature in the dark. F-actin (red) was stained with a fluorescent phalloidin and nuclei (blue) were stained with DAPI. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes at room temperature and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with Images were taken at a 60X magnification.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Hsp60 antibody [2E1/53] (ab5479)

  Immunofluorescence analysis of HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling Hsp60 (green) with ab5479 at 1/100 dilution or without overnight at 4 C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. 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. Images were taken at 60X magnification.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Hsp60 antibody [2E1/53] (ab5479)

  Immunofluorescence analysis of NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling Hsp60 (green) with ab5479 at 1/100 dilution or without (negative control) overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. 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. Images were taken at 60X magnification.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Hsp60 antibody [2E1/53] (ab5479)

  Immunofluorescence analysis of U-251 MG (formally U-373 MG) (human brain glioma cell line) cells labeling Hsp60 with ab5479 at 1/200 dilution overnight at 4°C, washed with PBS, followed by DyLight-488 conjugated secondary antibody.

  Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with ab5479. Heat Shock Protein 60 (Hsp60) staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Hsp60 antibody [2E1/53] (ab5479)

  Immunofluorescence analysis of HeLa (human epithelial cell line from cervix adenocarcinoma) and NIH/3T3 (mouse embryonic fibroblast cell line) cells labeling HSP60 (green) with ab5479 at 10ug/ml for at least 1 hour at room temperature. Cells were washed with PBS and incubated with a fluorescently labeled goat anti-mouse IgM secondary antibody at 1/400 dilution for 30 minutes at room temperature. F-Actin (red) was stained with DyLight 554 Phalloidin and nuclei (blue) were stained with Hoechst 33342 dye.

  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. Images were taken at 20X magnification.

• 

  Immunoprecipitation - Anti-Hsp60 antibody [2E1/53] (ab5479)

  Immunoprecipitation of Hsp60 was performed on HeLa (human epithelial cell line from cervix adenocarcinoma) cells. Antigen-antibody complexes were formed by incubating 500ug of whole cell lysate with 2ug of ab5479 overnight on a rocking platform at 4°C. The immune complexes were captured on 50ul Protein A/G Plus Agarose, washed extensively, and eluted with 5X Lane Marker Reducing Sample Buffer. Samples were resolved on a 4-20% Tris-HCl polyacrylamide gel, transferred to a PVDF membrane, and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with ab5479 at a concentration of 1ug/ml overnight rotating at 4°C, washed in TBST, and probed with a goat anti-mouse IgM secondary antibody at a dilution of 1:20,000 for at least 1 hour. Chemiluminescent detection performed using SuperSignal West Dura.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp60 antibody [2E1/53] (ab5479)

  Immunohistochemiscal analysis of both normal and cancer biopsies of deparaffinized human breast carcinoma tissues labeling Hsp60 with ab5479 at 1/20 dilution 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.

  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. 

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp60 antibody [2E1/53] (ab5479)

  Immunohistochemiscal analysis of both normal and cancer biopsies of deparaffinized human kidney tissue labeling Hsp60 with 1/100 dilution 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.

  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.