重组Anti-PFKFB3抗体[EPR12594] - BSA and Azide free (ab218121)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR12594] to PFKFB3 - BSA and Azide free
- Suitable for: Flow Cyt (Intra), IP, ICC/IF, WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-PFKFB3抗体[EPR12594] - BSA and Azide free
参阅全部 PFKFB3 一抗 -
描述
兔单克隆抗体[EPR12594] to PFKFB3 - BSA and Azide free -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), IP, ICC/IF, WB, IHC-Pmore 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: Jurkat and HeLa whole cell lysate (ab150035); Human melanoma tissue; HeLa and A431 cells, Mouse skin tissue lysate, Rat breast tissue lysate, AR42 and L6 whole cell lysates, HAP1 whole cell lysate, AR42J (rat pancreatic tumor epithelial cell) whole cell lysate, IP: Mouse skin tissue lysate, AR42J, whole cell lysate ICC: HeLa, A431 cells IHC: human melanoma tissue Flow: A431 (human epidermoid carcinoma) cells,
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常规说明
ab218121 is the carrier-free version of ab181861.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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. Do Not Freeze. -
存储溶液
pH: 7.2
Constituent: PBS -
无载体
是 -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR12594 -
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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Conjugation kits
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Isotype control
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Positive Controls
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab218121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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Flow Cyt (Intra) |
Use at an assay dependent concentration.
ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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IP |
Use at an assay dependent concentration.
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ICC/IF |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Detects a band of approximately 58 kDa (predicted molecular weight: 60 kDa).
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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说明 |
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Flow Cyt (Intra)
Use at an assay dependent concentration. ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
IP
Use at an assay dependent concentration. |
ICC/IF
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Detects a band of approximately 58 kDa (predicted molecular weight: 60 kDa). |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
靶标
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功能
Synthesis and degradation of fructose 2,6-bisphosphate. -
组织特异性
Ubiquitous. -
序列相似性
In the C-terminal section; belongs to the phosphoglycerate mutase family. -
翻译后修饰
Phosphorylation by AMPK stimulates activity. - Information by UniProt
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数据库链接
- Entrez Gene: 5209 Human
- Entrez Gene: 170768 Mouse
- Entrez Gene: 117276 Rat
- Omim: 605319 Human
- SwissProt: Q16875 Human
- SwissProt: A2AUP1 Mouse
- SwissProt: O35552 Rat
- Unigene: 195471 Human
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别名
- 6 phosphofructo 2 kinase/ fructose 2,6 bisphosphatase antibody
- 6 phosphofructo 2 kinase/fructose 2,6 biphosphatase 3 antibody
- 6-bisphosphatase antibody
see all
图片
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This WB data was generated using the same anti-PFKFB3 antibody clone, EPR12594, in a different buffer formulation (cat# ab181861).
Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: PFKFB3 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: Jurkat whole cell lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab181861 observed at 60 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab181861 was shown to specifically react with PFKFB3 in wild-type HAP1 cells as signal was lost in PFKFB3 knockout cells. Wild-type and PFKFB3 knockout samples were subjected to SDS-PAGE. Ab181861 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed A431 cells labeling PFKFB3 with ab181861 at 1/100 dilution, followed by Goat anti rabbit IgG (Alexa Fluor®555) at 1/200 dilution. Counter stained with Dapi (blue).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181861).
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All lanes : Anti-PFKFB3 antibody [EPR12594] (ab181861) at 1/20000 dilution
Lane 1 : Jurkat cell lysate
Lane 2 : HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution
Predicted band size: 60 kDa
Observed band size: 58 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST
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All lanes : Anti-PFKFB3 antibody [EPR12594] (ab181861) at 1/1000 dilution
Lane 1 : A431 (human epidermoid carcinoma epithelial cell) whole cell lysate
Lane 2 : Mouse skin tissue lysate
Lane 3 : Rat breast tissue lysate
Lane 4 : AR42J (rat pancreatic tumor epithelial cell) whole cell lysate
Lane 5 : L6 (rat skeletal muscle myoblast) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 60 kDa
Observed band size: 60 kDa
Exposure time: 15 secondsBlocking buffer: 5% NFDM/TBST
Exposure time: 15 seconds
This blot was developed using a high sensitivity ECL substrate.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181861).
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All lanes : Anti-PFKFB3 antibody [EPR12594] (ab181861) at 1/1000 dilution
Lane 1 : A431 (human epidermoid carcinoma epithelial cell), whole cell lysate
Lane 2 : bEnd.3 (mouse brain endothelial cell), whole cell lysate
Lane 3 : NIH/3T3 (mouse embryonic fibroblast), whole cell lysate
Lane 4 : 4T1 (mouse mammary gland carcinoma epithelial cell), whole cell lysate
Lane 5 : Undifferentiated 3T3-L1 (mouse embryonic fibroblast), whole cell lysate
Lane 6 : 3T3-L1 (mouse embryonic fibroblast) differentiated into adipocyte-like cells, whole cell lysate
Lane 7 : C2C12 (mouse myoblast), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 60 kDa
Observed band size: 60 kDaExposure time: Lane 1-2: 26 seconds, Lane 3-7: 48 seconds
Blocking buffer and concentration: 5% NFDM/TBST
Lane 3-7 were developed using a high sensitivity ECL substrate.
The expression level of PFKFB3 is upregulated during 3T3-L1 differentiation (PMID: 16306349).
The band at approximately 110 kDa is likely to be PFKFB3 dimer (PMID: 31889092).This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181861).
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This IHC data was generated using the same anti-PFKFB3 antibody clone, EPR12594, in a different buffer formulation (cat# ab181861).
Immunohistochemical analysis of paraffin-embedded human melanoma tissue labeling PFKFB3 with ab181861 at 1/50 dilution followed by prediluted HRP Polymer for Rabbit IgG. Counter stained with Hematoxylin.
Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
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Intracellular Flow Cytometry analysis of A431 (human epidermoid carcinoma) cells labeling PFKFB3 with purified ab181861 at 1/210 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181861).
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Western blot analysis of PFKFB3 in HeLa cell lysate immunoprecipitated using ab181861 at 1/50 dilution (Lane 1). Lane 2: Negative control.
Secondary antibody: Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1500 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181861).
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PFKFB3 was immunoprecipitated from 0.35 mg of Mouse skin tissue lysate with ab181861 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab181861 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.
Lane 1: Mouse skin tissue lysate 10 μg (Input).
Lane 2: ab181861 IP in Mouse skin tissue lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab181861 in Mouse skin tissue lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 180 seconds.
This blot was developed using a high sensitivity ECL substrate.
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PFKFB3 was immunoprecipitated from 0.35 mg of AR42J (rat pancreatic tumor epithelial cell) whole cell lysate with ab181861 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab181861 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.
Lane 1: AR42J (rat pancreatic tumor epithelial cell) whole cell lysate 10 μg (Input).
Lane 2: ab181861 IP in AR42J whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab181861 in AR42J whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 180 seconds.
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Immunofluorescent analysis of acetone-fixed HeLa cells labeling PFKFB3 with ab181861 at 1/100 dilution, followed by Goat anti rabbit IgG (Alexa Fluor®555) at 1/200 dilution. Counter stained with Dapi (blue).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181861).
实验方案
数据表及文件
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Datasheet download
Certificate of Compliance
文献 (4)
ab218121 被引用在 4 文献中.
- Tang CJ et al. Metformin prevents PFKFB3-related aerobic glycolysis from enhancing collagen synthesis in lung fibroblasts by regulating AMPK/mTOR pathway. Exp Ther Med 21:581 (2021). PubMed: 33850553
- Zhang J et al. Protein kinase D3 promotes gastric cancer development through p65/6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 activation of glycolysis. Exp Cell Res 380:188-197 (2019). PubMed: 31026442
- Li Z et al. The immunoregulatory protein B7-H3 promotes aerobic glycolysis in oral squamous carcinoma via PI3K/Akt/mTOR pathway. J Cancer 10:5770-5784 (2019). PubMed: 31737114
- Jiang H et al. PFKFB3-Driven Macrophage Glycolytic Metabolism Is a Crucial Component of Innate Antiviral Defense. J Immunol 197:2880-90 (2016). PubMed: 27566823