重组Anti-Hexokinase 1抗体[EPR10134(B)] - Mitochondrial Outer膜Marker (ab150423)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR10134(B)] to Hexokinase 1 - Mitochondrial Outer Membrane Marker
- Suitable for: WB, IHC-P, Flow Cyt (Intra), ICC/IF, mIHC
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-Hexokinase 1抗体[EPR10134(B)] - Mitochondrial Outer膜Marker
参阅全部 Hexokinase 1 一抗 -
描述
兔单克隆抗体[EPR10134(B)] to Hexokinase 1 - Mitochondrial Outer膜Marker -
宿主
Rabbit -
经测试应用
适用于: WB, IHC-P, Flow Cyt (Intra), ICC/IF, mIHCmore details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Synthetic peptide corresponding to Human Hexokinase 1 aa 100-200 (internal sequence).
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阳性对照
- WB: HEK-293T and MCF7 cells, human mouse and rat brain lysates; IHC: Human, mouse and rat kidney tissue; ICC/IF: MCF7 lysate; Flow Cyt (intra): K-562 cells. mIHC-P: Human kidney tissue.
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常规说明
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
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. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR10134(B) -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
- Alexa Fluor® 488 Anti-Hexokinase 1 antibody [EPR10134(B)] - Mitochondrial Outer Membrane Marker (ab184818)
- Alexa Fluor® 647 Anti-Hexokinase 1 antibody [EPR10134(B)] - Mitochondrial Outer Membrane Marker (ab197864)
- Anti-Hexokinase 1 antibody [EPR10134(B)] - BSA and Azide free (ab233837)
- Alexa Fluor® 568 Anti-Hexokinase 1 antibody [EPR10134(B)] - Mitochondrial Outer Membrane Marker (ab312784)
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab150423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 - 1/10000. Predicted molecular weight: 102 kDa.
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IHC-P |
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Flow Cyt (Intra) |
1/20.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ICC/IF |
1/50.
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mIHC |
1/250.
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说明 |
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WB
1/1000 - 1/10000. Predicted molecular weight: 102 kDa. |
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Flow Cyt (Intra)
1/20. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ICC/IF
1/50. |
mIHC
1/250. |
靶标
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组织特异性
Isoform 2 is erythrocyte specific. Isoform 3 and isoform 4 are testis-specific. -
通路
Carbohydrate metabolism; hexose metabolism. -
疾病相关
Hexokinase deficiency
Neuropathy, hereditary motor and sensory, Russe type -
序列相似性
Belongs to the hexokinase family.
Contains 2 hexokinase domains. -
结构域
The N- and C-terminal halves of this hexokinase show extensive sequence similarity to each other. The catalytic activity is associated with the C-terminus while regulatory function is associated with the N-terminus. Each domain can bind a single glucose and Gluc-6-P molecule. -
细胞定位
Mitochondrion outer membrane. Its hydrophobic N-terminal sequence may be involved in membrane binding. - Information by UniProt
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数据库链接
- Entrez Gene: 3098 Human
- Entrez Gene: 15275 Mouse
- Entrez Gene: 25058 Rat
- Omim: 142600 Human
- SwissProt: P19367 Human
- SwissProt: P17710 Mouse
- SwissProt: P05708 Rat
- Unigene: 370365 Human
see all -
别名
- BB404130 antibody
- Brain form hexokinase antibody
- dea antibody
see all
图片
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Fluorescence multiplex immunohistochemical analysis of the human kidney (Formalin/PFA-fixed paraffin-embedded sections).
Panel A: merged staining of anti-Hexokinase 1 (ab150423, green; Opal™690), anti-Angiotensin Converting Enzyme 1 (ab254222, gray; Opal™520) and anti-Aquaporin 2 (ab199975, red; Opal™570) on human kidney. Panel B: anti-Aquaporin 2 stained on collecting tubules. Panel C: anti-Angiotensin Converting Enzyme 1 stained on proximal tubules. Panel D: anti-Hexokinase 1 stained on distal tubules and collecting tubules. Opal Polymer HRP Ms + Rb was used as a secondary antibody.
The section was incubated in three rounds of staining: in the order of ab150423 at 1/250 dilution (4.224 μg/ml), ab254222 at 1/4000 dilution (0.141 μg/ml) and ab199975 at 1/4000 dilution (0.152 μg/ml) for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. DAPI (blue) was used as a nuclear counter stain.
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Fluorescence multiplex immunohistochemical analysis of paraffin-embedded Human kidney tissue.
Panel A: Merged staining of anti-Hexokinase 1 (gray; Opal™690), anti-Angiotensin Converting Enzyme 1 (green; Opal™520) and anti-Tissue Factor (red; Opal™570) on human kidney.
Panel B: Anti-Tissue Factor stained on renal glomeruli.
Panel C: Anti-Angiotensin Converting Enzyme 1 stained on proximal tubules.
Panel D: Anti-Hexokinase 1 stained on distal tubules.The section was incubated in three rounds of staining: in the order of ab150423, ab254222, and ab252918 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins. Counterstained with DAPI.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat kidney tissue sections labeling Hexokinase 1 with purified ab150423 at 1/50 dilution (4.14 µg/mL). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse kidney tissue sections labeling Hexokinase 1 with purified ab150423 at 1/50 dilution (4.14 µg/mL). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human kidney tissue sections labeling Hexokinase 1 with purified ab150423 at 1/50 dilution (4.14 µg/mL). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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All lanes : Anti-Hexokinase 1 antibody [EPR10134(B)] - Mitochondrial Outer Membrane Marker (ab150423) at 1/1000 dilution (Purified)
Lane 1 : Human brain lysate
Lane 2 : Mouse brain lysate
Lane 3 : Rat brain lysate
Lane 4 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate
Lane 5 : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 102 kDa -
Intracellular Flow Cytometry analysis of K-562 (Human chronic myelogenous leukemia lymphoblast) cells labeling Hexokinase 1 with Purified ab150423 at 1/20 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488 ,ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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Immunocytochemistry analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling Hexokinase 1 with Purified ab150423 at 1:50 dilution (4.1 µg/ml). Cells were fixed in 100% Methanol and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488,ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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All lanes : Anti-Hexokinase 1 antibody [EPR10134(B)] - Mitochondrial Outer Membrane Marker (ab150423) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : HK1 knockout HeLa cell lysate
Lane 3 : MCF7 cell lysate
Lane 4 : HEPG2 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 102 kDa
Observed band size: 102 kDaLanes 1-4: Merged signal (red and green). Green - ab150423 observed at 102 kDa. Red - loading control, ab8245 observed at 37 kDa.
ab150423 Anti-Hexokinase 1 antibody [EPR10134(B)] - Mitochondrial was shown to specifically react with Hexokinase 1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab267279 (knockout cell lysate ab257161) was used. Wild-type and Hexokinase 1 knockout samples were subjected to SDS-PAGE. ab150423 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (12)
ab150423 被引用在 12 文献中.
- Wang M et al. Ceria nanoparticles ameliorate renal fibrosis by modulating the balance between oxidative phosphorylation and aerobic glycolysis. J Nanobiotechnology 20:3 (2022). PubMed: 34983531
- Wang D et al. Lactate oxidative phosphorylation by annulus fibrosus cells: evidence for lactate-dependent metabolic symbiosis in intervertebral discs. Arthritis Res Ther 23:145 (2021). PubMed: 34020698
- Shi L et al. Spaceflight and simulated microgravity suppresses macrophage development via altered RAS/ERK/NF?B and metabolic pathways. Cell Mol Immunol 18:1489-1502 (2021). PubMed: 31900461
- Bennett NK et al. Defining the ATPome reveals cross-optimization of metabolic pathways. Nat Commun 11:4319 (2020). PubMed: 32859923
- Subramani J et al. Thioredoxin protects mitochondrial structure, function and biogenesis in myocardial ischemia-reperfusion via redox-dependent activation of AKT-CREB- PGC1a pathway in aged mice. Aging (Albany NY) 12:19809-19827 (2020). PubMed: 33049718