重组Anti-PFKFB3抗体[EPR12594] (ab181861)
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
- Suitable for: WB, IP, ICC/IF, IHC-P, Flow Cyt
- Knockout validated
- Reacts with: Human
概述
-
产品名称
Anti-PFKFB3抗体[EPR12594]
参阅全部 PFKFB3 一抗 -
描述
兔单克隆抗体[EPR12594] to PFKFB3 -
宿主
Rabbit -
Tested Applications & Species
Application Species Flow Cyt HumanICC/IF HumanIHC-P HumanIP HumanWB Human -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
-
阳性对照
- Jurkat and HeLa whole cell lysate (ab150035); Human melanoma tissue; HeLa and A431 cells.
-
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.
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 the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.
One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.
Learn more here.
性能
-
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
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
-
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
-
Alternative Versions
-
Compatible Secondaries
-
Conjugation kits
-
Isotype control
-
Positive Controls
-
Recombinant Protein
-
Related Products
应用
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab181861 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
应用 | Species |
---|---|
Flow Cyt |
Human
|
ICC/IF |
Human
|
IHC-P |
Human
|
IP |
Human
|
WB |
Human
|
All applications |
Mouse
Rat
|
应用 | Ab评论 | 说明 |
---|---|---|
WB | (4) |
1/1000 - 1/10000. Detects a band of approximately 58 kDa (predicted molecular weight: 60 kDa).
|
IP |
1/50.
|
|
ICC/IF |
1/100.
|
|
IHC-P |
1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
|
|
Flow Cyt |
Use at an assay dependent concentration.
|
说明 |
---|
WB
1/1000 - 1/10000. Detects a band of approximately 58 kDa (predicted molecular weight: 60 kDa). |
IP
1/50. |
ICC/IF
1/100. |
IHC-P
1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Flow Cyt
Use at an assay dependent concentration. |
靶标
-
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
-
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
see all -
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
-
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.
-
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).
-
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
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PFKFB3 antibody [EPR12594] (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.
-
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.
-
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.
-
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).
实验方案
数据表及文件
-
SDS download
-
Datasheet download
Certificate of Compliance
文献 (17)
ab181861 被引用在 17 文献中.
- Piers TM et al. A locked immunometabolic switch underlies TREM2 R47H loss of function in human iPSC-derived microglia. FASEB J 34:2436-2450 (2020). PubMed: 31907987
- Liu J et al. Long noncoding RNA AGPG regulates PFKFB3-mediated tumor glycolytic reprogramming. Nat Commun 11:1507 (2020). PubMed: 32198345
- Zhang J et al. Dual inhibition of PFKFB3 and VEGF normalizes tumor vasculature, reduces lactate production, and improves chemotherapy in glioblastoma: insights from protein expression profiling and MRI. Theranostics 10:7245-7259 (2020). PubMed: 32641990
- Pan H et al. RNA binding protein HuR regulates extracellular matrix gene expression and pH homeostasis independent of controlling HIF-1a signaling in nucleus pulposus cells. Matrix Biol 77:23-40 (2019). PubMed: 30092282
- Wang L et al. Ablation of endothelial Pfkfb3 protects mice from acute lung injury in LPS-induced endotoxemia. Pharmacol Res 146:104292 (2019). PubMed: 31167111