Recombinant Anti-PPP2R5E antibody [EPR17147] - C-terminal (ab198500)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR17147] to PPP2R5E - C-terminal
- Suitable for: WB, IP, ICC/IF, IHC-P, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
-
Product name
Anti-PPP2R5E antibody [EPR17147] - C-terminal
See all PPP2R5E primary antibodies -
Description
Rabbit monoclonal [EPR17147] to PPP2R5E - C-terminal -
Host species
Rabbit -
Tested applications
Suitable for: WB, IP, ICC/IF, IHC-P, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
-
Positive control
- WB: HeLa, HEK-293T and Daudi cell lysates; Human skeletal muscle lysates. IHC: Human cervix carcinoma and bladder transitional cell carcinoma tissues. ICC/IF: HeLa cells. Flow Cyt (intra): HeLa cell lysate. IP: HeLa 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: 40% Glycerol (glycerin, glycerine), 59% PBS, 0.05% BSA -
Concentration information loading...
-
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR17147 -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
KO cell lines
-
KO cell lysates
-
Positive Controls
-
Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab198500 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 |
1/5000. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa).
|
|
IP |
1/75.
|
|
ICC/IF |
1/500.
|
|
IHC-P |
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
|
|
Flow Cyt (Intra) |
1/2500.
|
Notes |
---|
WB
1/5000. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa). |
IP
1/75. |
ICC/IF
1/500. |
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Flow Cyt (Intra)
1/2500. |
Target
-
Function
The B regulatory subunit might modulate substrate selectivity and catalytic activity, and also might direct the localization of the catalytic enzyme to a particular subcellular compartment. -
Sequence similarities
Belongs to the phosphatase 2A regulatory subunit B56 family. -
Post-translational
modificationsPhosphorylated on serine residues. -
Cellular localization
Cytoplasm. - Information by UniProt
-
Database links
- Entrez Gene: 5529 Human
- Entrez Gene: 26932 Mouse
- Entrez Gene: 299147 Rat
- Omim: 601647 Human
- SwissProt: Q16537 Human
- SwissProt: Q61151 Mouse
- Unigene: 334868 Human
- Unigene: 259626 Mouse
see all -
Alternative names
- 2A5E_HUMAN antibody
- Epsilon isoform of regulatory subunit B56 protein phosphatase 2A antibody
- PP2A B subunit B' epsilon antibody
see all
Images
-
All lanes : Anti-PPP2R5E antibody [EPR17147] - C-terminal (ab198500) at 1/5000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : PPP2R5E knockout HeLa cell lysate
Lane 3 : HEK-293T cell lysate
Lane 4 : Daudi cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 55 kDaLanes 1-4: Merged signal (red and green). Green - ab198500 observed at 55 kDa. Red - loading control ab8245 observed at 37 kDa.
ab198500 Recombinant Anti-PPP2R5E antibody [EPR17147] - C-terminal was shown to specifically react with PPP2R5E in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265637 (knockout cell lysate ab258135) was used. Wild-type and PPP2R5E knockout samples were subjected to SDS-PAGE. ab198500 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 5000 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 20000 dilution for 1 hour at room temperature before imaging.
-
Immunohistochemical analysis of paraffin-embedded Human cervix carcinoma tissue labeling PPP2R5E with ab198500 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on Human cervix carcinoma tissue isobserved. Counter-stained with Hematoxylin.
Negative control: Using PBS instead of primary antibody.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MCF-7 (Human breast carcinoma) cells labeling PPP2R5E with ab198500 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Cytoplasm staining on MCF7 cell line is observed. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows;
1. ab198500 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
2. ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution. -
Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling PPP2R5E (red) with purified ab198500 at a 1/2500 dilution (1ug/mL). Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti rabbit IgG (Alexa Fluorr® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (Black) (ab172730). Blue (unlabeled control) - Cell without incubation with primary antibody and secondary antibody (Blue).
-
All lanes : Anti-PPP2R5E antibody [EPR17147] - C-terminal (ab198500) at 1/5000 dilution
Lane 1 : HeLa cell lysate at 10 µg
Lane 2 : 293 cell lysate at 10 µg
Lane 3 : Human skeletal muscle lysate at 20 µg
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution
Developed using the ECL technique.
Predicted band size: 55 kDa
Observed band size: 55 kDa
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
-
PPP2R5E was immunoprecipitated from 1mg of HeLa (Human cervix adenocarcinoma) whole cell extract with ab198500 at 1/175 dilution. Western blot was performed from the immunoprecipitate using ab198500 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: HeLa whole cell extract 10µg (Input).
Lane 2: HeLa whole cell extract
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab198500 in HeLa whole cell extract.
Blocking and dilution buffer and concentration: 5% NFDM/TBST. -
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human cervix adenocarcinoma) cells labeling PPP2R5E with ab198500 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Cytoplasm staining on HeLa cell line is observed. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows;
1. ab198500 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
2. ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution. -
Immunohistochemical analysis of paraffin-embedded Human transitional cell carcinoma of bladder tissue labeling PPP2R5E with ab198500 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on Human transitional cell carcinoma of bladder tissue isobserved. Counter-stained with Hematoxylin.
Negative control: Using PBS instead of primary antibody.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Protocols
Datasheets and documents
-
SDS download
-
Datasheet download
Certificate of Compliance
References (0)
ab198500 has not yet been referenced specifically in any publications.