重组Anti-Hsp60抗体[EPR18245-93] (ab190828)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18245-93] to Hsp60
- Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-Hsp60抗体[EPR18245-93]
参阅全部 Hsp60 一抗 -
描述
兔单克隆抗体[EPR18245-93] to Hsp60 -
宿主
Rabbit -
经测试应用
适用于: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)more details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: Human brain, fetal heart, fetal kidney and fetal spleen lysates; HeLa, NIH/3T3, Neuro-2a and C6 cell lysates; Mouse heart and spleen lysates; Rat brain, heart, spleen and liver lysates. IHC-P: Human, mouse and rat liver tissues. ICC/IF: HeLa and NIH/3T3 cells. Flow Cyt (intra): NIH/3T3 cells. IP: NIH/3T3 cell lysate.
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常规说明
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
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 long term. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 0.05% BSA, 40% Glycerol (glycerin, glycerine), PBS -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR18245-93 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab190828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。
应用 | Ab评论 | 说明 |
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WB | (4) |
1/1000. Detects a band of approximately 61 kDa (predicted molecular weight: 61 kDa).
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IHC-P |
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
1/200.
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IP |
1/30.
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Flow Cyt (Intra) |
1/500.
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说明 |
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WB
1/1000. Detects a band of approximately 61 kDa (predicted molecular weight: 61 kDa). |
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/200. |
IP
1/30. |
Flow Cyt (Intra)
1/500. |
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: 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
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 [EPR18245-93] (ab190828) at 1/1000 dilution
Lane 1 : Human brain lysate
Lane 2 : Human fetal heart lysate
Lane 3 : Human fetal kidney lysate
Lane 4 : Human fetal spleen lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution
Developed using the ECL technique.
Predicted band size: 61 kDa
Observed band size: 61 kDaExposure time : Lanes 1,2 and 3: 3 minutes; Lane 4:1 minute.
Blocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-Hsp60 antibody [EPR18245-93] (ab190828) at 1/5000 dilution
Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate
Lane 2 : NIH/3T3 (mouse embyro fibroblast cell line) cell lysate
Lane 3 : Neuro-2a (mouse neuroblastoma cell line) cell lysate
Lane 4 : C6 (rat glial tumor cell line) cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 61 kDa
Observed band size: 61 kDaExposure time : Lanes 1,2 and 3: 30 seconds; Lane 4:15 seconds.
Blocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-Hsp60 antibody [EPR18245-93] (ab190828) at 1/1000 dilution
Lane 1 : Mouse heart lysate
Lane 2 : Mouse spleen lysate
Lane 3 : Rat brain lysate
Lane 4 : Rat heart lysate
Lane 5 : Rat spleen lysate
Lane 6 : Rat liver lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 61 kDa
Observed band size: 61 kDaExposure time : Lane 1: 30 seconds; Lane 2: 1 minute; Lanes 3,4 and 5: 10 seconds; Lane 6: 5 seconds.
Blocking/Dilution buffer: 5% NFDM/TBST.
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Immunohistochemical analysis of paraffin-embedded human liver tissue labeling Hsp60 with ab190828 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Granular and cytoplasmic staining on human liver (PMID: 18548335). Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling Hsp60 with ab190828 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Granular and cytoplasmic staining on mouse liver (PMID: 18548335). Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded rat liver tissue labeling Hsp60 with ab190828 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Granular and cytoplasmic staining on rat liver (PMID: 18548335). Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling Hsp60 with ab190828 at 1/200 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HeLa cells.
The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling Hsp60 with ab190828 at 1/200 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HeLa cells.
The nuclear counter stain is DAPI (blue). COX IV is detected with Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker (ab33985) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) (red) at 1/1000 dilution.
The negative controls are as follows:-
-ve control 1: ab190828 at 1/200 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution.
-ve control 2: Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker (ab33985) at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (mouse embyro fibroblast cell line) cells labeling Hsp60 with ab190828 at 1/200 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on NIH/3T3 cells.
The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized NIH/3T3 (mouse embyro fibroblast cell line) cell line labeling Hsp60 with ab190828 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluorr® 488) at 1/2000 dilution was used as the secondary antibody.
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Hsp60 was immunoprecipitated from 0.35 mg of NIH/3T3 (mouse embyro fibroblast cell line) cell lysate with ab190828 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab190828 at 1/5000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: NIH/3T3 cell lysate 10 µg (Input).
Lane 2: ab190828 IP in NIH/3T3 cell lysate .
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab190828 in NIH/3T3 cell lysate .
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 1 second.
实验方案
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (9)
ab190828 被引用在 9 文献中.
- D'Acunzo P et al. Isolation of mitochondria-derived mitovesicles and subpopulations of microvesicles and exosomes from brain tissues. Nat Protoc 17:2517-2549 (2022). PubMed: 35962195
- Ren J et al. Targeted activation of GPER enhances the efficacy of venetoclax by boosting leukemic pyroptosis and CD8+ T cell immune function in acute myeloid leukemia. Cell Death Dis 13:915 (2022). PubMed: 36316313
- Feng Y et al. Dl-3-n-Butylphthalide Alleviates Demyelination and Improves Cognitive Function by Promoting Mitochondrial Dynamics in White Matter Lesions. Front Aging Neurosci 13:632374 (2021). PubMed: 33762923
- Xi H et al. FOXG1 improves mitochondrial function and promotes the progression of nasopharyngeal carcinoma. Mol Med Rep 24:N/A (2021). PubMed: 34278485
- Zhang L et al. Transferrin receptor-mediated reactive oxygen species promotes ferroptosis of KGN cells via regulating NADPH oxidase 1/PTEN induced kinase 1/acyl-CoA synthetase long chain family member 4 signaling. Bioengineered 12:4983-4994 (2021). PubMed: 34369274
- Gao X et al. ATF5, a putative therapeutic target for the mitochondrial DNA 3243A > G mutation-related disease. Cell Death Dis 12:701 (2021). PubMed: 34262025
- Casellas-Díaz S et al. Mfn2 localization in the ER is necessary for its bioenergetic function and neuritic development. EMBO Rep 22:e51954 (2021). PubMed: 34296790
- Cao LL et al. Downregulating expression of OPTN elevates neuroinflammation via AIM2 inflammasome- and RIPK1-activating mechanisms in APP/PS1 transgenic mice. J Neuroinflammation 18:281 (2021). PubMed: 34861878
- Zhang M et al. NAD+ repletion inhibits the endothelial-to-mesenchymal transition induced by TGF-ß in endothelial cells through improving mitochondrial unfolded protein response. Int J Biochem Cell Biol 117:105635 (2019). PubMed: 31626975