Recombinant Anti-PFKM + PFKP antibody [EPR17314] (ab204131)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR17314] to PFKM + PFKP
- Suitable for: WB, ICC/IF, IP, Flow Cyt (Intra)
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
-
Product name
Anti-PFKM + PFKP antibody [EPR17314] -
Description
Rabbit monoclonal [EPR17314] to PFKM + PFKP -
Host species
Rabbit -
Tested applications
Suitable for: WB, ICC/IF, IP, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
This product was produced with the following immunogens:
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers. -
Positive control
- WB: MCF-7, HeLa, Raji, Jurkat and NIH/3T3 whole cell lsates; Human fetal brain and fetal kidney lysates; Mouse and rat kidney and spleen lysates. ICC/IF: HeLa and MCF-7 cells. Flow Cyt (intra): Jurkat and MCF-7 cells. IP: MCF-7 whole cell lysate.
-
General notes
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.
Properties
-
Form
Liquid -
Storage instructions
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. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
-
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR17314 -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Conjugation kits
-
Isotype control
-
Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab204131 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
---|---|---|
WB |
1/2000. Detects a band of approximately 86 kDa (predicted molecular weight: 86 kDa).
|
|
ICC/IF |
1/250.
|
|
IP |
1/80.
|
|
Flow Cyt (Intra) |
1/250.
|
Notes |
---|
WB
1/2000. Detects a band of approximately 86 kDa (predicted molecular weight: 86 kDa). |
ICC/IF
1/250. |
IP
1/80. |
Flow Cyt (Intra)
1/250. |
Images
-
All lanes : Anti-PFKM + PFKP antibody [EPR17314] (ab204131) at 1/20000 dilution
Lane 1 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
Lane 2 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
Lane 3 : Raji (Human Burkitt's lymphoma cell line) whole cell lysate
Lane 4 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Predicted band size: 86 kDa
Observed band size: 86 kDa
Exposure time: 5 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
-
All lanes : Anti-PFKM + PFKP antibody [EPR17314] (ab204131) at 1/2000 dilution
Lane 1 : Human fetal brain lysate
Lane 2 : Human fetal kidney lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/50000 dilution
Predicted band size: 86 kDa
Observed band size: 86 kDa
Exposure time: 5 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
-
All lanes : Anti-PFKM + PFKP antibody [EPR17314] (ab204131) at 1/2000 dilution
Lane 1 : Mouse kidney lysate
Lane 2 : Mouse spleen lysate
Lane 3 : Rat kidney lysate
Lane 4 : Rat spleen lysate
Lane 5 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Predicted band size: 86 kDa
Observed band size: 86 kDa
Exposure time: 5 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
-
All lanes : Anti-PFKM + PFKP antibody [EPR17314] (ab204131) at 1/1000 dilution
Lane 1 :Recombinant Human PFKP protein (ab132823)
Lane 2 : DDK tagged Recombinant Human PFKM protein (Full length, 85 KDa)
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 86 kDaBlocking/diluting buffer and concentration: 5% NFDM/TBST
Exposure time: Lane1: 10 seconds
Lane 2: 1 seconds -
Immunofluorescent analysis of 100% methanl-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling PFKP with ab204131 at 1/250 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 cell line. The nuclear counter stain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: ab204131 at 1/250 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution.
-ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution. -
Immunofluorescent analysis of 100% methanl-fixed, 0.1% Triton X-100 permeabilized MCF7 (Human breast adenocarcinoma cell line) cells labeling PFKP with ab204131 at 1/250 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 MCF7 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: ab204131 at 1/250 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution.
-ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution. -
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling PFKP with ab204131 at 1/250 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/500 dilution was used as the secondary antibody.
-
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed MCF7 (Human breast adenocarcinoma cell line) cells labeling PFKP with ab204131 at 1/250 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/500 dilution was used as the secondary antibody.
-
PFKP was immunoprecipitated from 1mg of MCF7 (Human breast adenocarcinoma cell line) whole cell lysate with ab204131 at 1/80 dilution.
Western blot was performed from the immunoprecipitate using ab204131 at 1/1000 dilution.
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.
Lane 1: MCF7 whole cell lysate 10ug (Input).
Lane 2: ab204131 IP in MCF7 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab204131 in MCF7 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 10 seconds.
Protocols
Datasheets and documents
-
SDS download
-
Datasheet download
Certificate of Compliance
References (11)
ab204131 has been referenced in 11 publications.
- 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
- Chen S et al. Epigallocatechin‑3‑gallate hinders metabolic coupling to suppress colorectal cancer malignancy through targeting aerobic glycolysis in cancer‑associated fibroblasts. Int J Oncol 60:N/A (2022). PubMed: 35029285
- Chen X et al. Intracellular galectin-3 is a lipopolysaccharide sensor that promotes glycolysis through mTORC1 activation. Nat Commun 13:7578 (2022). PubMed: 36481721
- 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
- Shen J et al. PFKP is highly expressed in lung cancer and regulates glucose metabolism. Cell Oncol (Dordr) N/A:N/A (2020). PubMed: 32219704