Anti-Hsp60抗体[4B9/89] (ab5478)
Key features and details
- Mouse monoclonal [4B9/89] to Hsp60
- Suitable for: ICC/IF, IP, ICC, WB
- Reacts with: Mouse, Rat, Hamster, Human, Non human primates
- Isotype: IgG2a
概述
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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: ICC/IF, IP, ICC, WBmore details -
种属反应性
与反应: Mouse, Rat, Hamster, Human, Non human primates -
免疫原
Other Immunogen Type corresponding to Human Hsp60. Human placental Hsp60.
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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. -
阳性对照
- WB: human blood. ICC: B-SC-1 cells.
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常规说明
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
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, as well as customer reviews and Q&As.
性能
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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 -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
4B9/89 -
同种型
IgG2a -
研究领域
相关产品
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Compatible Secondaries
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Isotype control
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Recombinant Protein
应用
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.
应用 | Ab评论 | 说明 |
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ICC/IF | 1/50. | |
IP | Use a concentration of 2 µg/ml. | |
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. |
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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. |
靶标
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功能
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. -
疾病相关
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. -
序列相似性
Belongs to the chaperonin (HSP60) family. -
细胞定位
Mitochondrion matrix. - Information by UniProt
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数据库链接
- Entrez Gene: 101844216 Hamster
- Entrez Gene: 3329 Human
- Entrez Gene: 15510 Mouse
- Entrez Gene: 63868 Rat
- Omim: 118190 Human
- SwissProt: P10809 Human
- SwissProt: P63038 Mouse
- SwissProt: P63039 Rat
see all -
别名
- 60 kDa chaperonin antibody
- 60 kDa heat shock protein, mitochondrial antibody
- CH60_HUMAN antibody
see all
图片
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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 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.
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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.
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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.
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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.
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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.
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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.
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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.
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Immunolocalization of Hsp 60 in human endothelial cells using ab5478.
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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.
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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.
实验方案
文献 (3)
ab5478 被引用在 3 文献中.
- Cheng Y et al. Expression and location of HSP60 and HSP10 in the heart tissue of heat-stressed rats. Exp Ther Med 12:2759-2765 (2016). IHC ; Rat . PubMed: 27698781
- Dai X et al. Requirement for integrin-linked kinase in neural crest migration and differentiation and outflow tract morphogenesis. BMC Biol 11:107 (2013). IHC-P . PubMed: 24131868
- 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 . PubMed: 17272814