Recombinant Anti-PPP2R5D antibody [EPR15617] (ab188323)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR15617] to PPP2R5D
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IP
- Knockout validated
- Reacts with: Rat, Human
Related conjugates and formulations
Overview
-
Product name
Anti-PPP2R5D antibody [EPR15617]
See all PPP2R5D primary antibodies -
Description
Rabbit monoclonal [EPR15617] to PPP2R5D -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IPmore details -
Species reactivity
Reacts with: Rat, Human
Predicted to work with: Mouse -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
-
Positive control
- WB: A431, MCF7, C6, HepG2, Jurkat, HeLa, 293T cell lysate. IHC-P: Human brain tissue. IP: HEK293 cells. Flow Cyt (intra): HeLa cells.
-
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
EPR15617 -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Isotype control
-
KO cell lines
-
KO cell lysates
-
Positive Controls
-
Recombinant Protein
-
Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab188323 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 |
---|---|---|
Flow Cyt (Intra) |
1/40.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
|
WB |
1/10000 - 1/50000. Detects a band of approximately 58, 66, 70 kDa (predicted molecular weight: 70 kDa).
|
|
IHC-P |
1/50 - 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
|
|
ICC/IF |
1/250.
|
|
IP |
1/40 - 1/60.
|
Notes |
---|
Flow Cyt (Intra)
1/40. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/10000 - 1/50000. Detects a band of approximately 58, 66, 70 kDa (predicted molecular weight: 70 kDa). |
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/250. |
IP
1/40 - 1/60. |
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. -
Tissue specificity
Isoform Delta-2 is widely expressed. Isoform Delta-1 is highly expressed in brain. -
Sequence similarities
Belongs to the phosphatase 2A regulatory subunit B56 family. -
Cellular localization
Cytoplasm. Nucleus. Nuclear in interphase, nuclear during mitosis. - Information by UniProt
-
Database links
- Entrez Gene: 5528 Human
- Entrez Gene: 21770 Mouse
- Entrez Gene: 363193 Rat
- Omim: 601646 Human
- SwissProt: Q14738 Human
- Unigene: 533308 Human
-
Alternative names
- 2A5D_HUMAN antibody
- B'delta antibody
- B56D antibody
see all
Images
-
All lanes : Anti-PPP2R5D antibody [EPR15617] (ab188323) at 1/10000 dilution
Lane 1 : Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 2 : PPP2R5D knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 4 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 70 kDa
Observed band size: 60-65 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab188323 observed at 60-65 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab188323 was shown to react with PPP2R5D in wild-type A431 cells in western blot with loss of signal observed in PPP2R5D knockout sample. Wild-type and PPP2R5D knockout A431 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab188323 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 10000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 hour at room temperature before imaging.
-
Immunohistochemical analysis of paraffin-embedded Human brain tissue labeling PPP2R5D using ab188323 at 1/100 dilution. A Ready to use HRP Polymer for Rabbit IgG (prediluted) was used as secondary. Counterstain: Hematoxylin.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
All lanes : Anti-PPP2R5D antibody [EPR15617] (ab188323) at 1/100000 dilution
Lane 1 : C6 cell lysate
Lane 2 : HepG2 cell lysate
Lane 3 : 293T cell lysate
Lane 4 : HeLa cell lysate
Lane 5 : Jurkat cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 70 kDa
Observed band size: 70 kDaBased on the sequence analysis, ab188323 recognizes 3 isoforms with the predicted MWs of 58KDa, 66KDa and 70KDa, respectively.
-
Immunoprecipitation analysis of 293T cell lysate labeling PPP2R5D using ab188323 at 1/50 dilution. A Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1500 was used as secondary antibody.
-
Immunofluorescence analysis of HepG2 cells labeling PPP2R5D using ab188323 at 1/250 dilution. A Goat anti rabbit IgG (Alexa Fluor488) at 1/200 dilution was used as secondary antibody. Cells were fixed with 4% Paraformaldehyde. Counterstain: DAPI.
-
Intracellular Flow Cytometry analysis of HeLa cells labeling PPP2R5D using ab188323 at 1/40 dilution (pink). A Goat anti rabbit IgG (FITC) at 1/150 dilution was used as secondary antibody. Cells were fixed with 2% paraformaldehyde. Isotype control: Rabbit monoclonal IgG (green).
Protocols
Datasheets and documents
-
SDS download
-
Datasheet download
Certificate of Compliance
References (6)
ab188323 has been referenced in 6 publications.
- Papke CM et al. A disorder-related variant (E420K) of a PP2A-regulatory subunit (PPP2R5D) causes constitutively active AKT-mTOR signaling and uncoordinated cell growth. J Biol Chem 296:100313 (2021). PubMed: 33482199
- Jong CJ et al. Reduction of protein phosphatase 2A (PP2A) complexity reveals cellular functions and dephosphorylation motifs of the PP2A/B'd holoenzyme. J Biol Chem 295:5654-5668 (2020). PubMed: 32156701
- Marelli S et al. Antagonism of PP2A is an independent and conserved function of HIV-1 Vif and causes cell cycle arrest. Elife 9:N/A (2020). PubMed: 32292164
- Naamati A et al. Functional proteomic atlas of HIV infection in primary human CD4+ T cells. Elife 8:N/A (2019). PubMed: 30857592
- Evans EL et al. HIV-1 Vif's Capacity To Manipulate the Cell Cycle Is Species Specific. J Virol 92:N/A (2018). PubMed: 29321323
- Greenwood EJ et al. Temporal proteomic analysis of HIV infection reveals remodelling of the host phosphoproteome by lentiviral Vif variants. Elife 5:N/A (2016). PubMed: 27690223