Anti-PIP2 antibody [2C11] (ab11039)
Key features and details
- Mouse monoclonal [2C11] to PIP2
- Suitable for: IHC-P, ICC/IF, ELISA, Neutralising
- Reacts with: Species independent
- Isotype: IgM
Overview
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Product name
Anti-PIP2 antibody [2C11] -
Description
Mouse monoclonal [2C11] to PIP2 -
Host species
Mouse -
Tested applications
Suitable for: IHC-P, ICC/IF, ELISA, Neutralisingmore details -
Species reactivity
Reacts with: Species independent -
Immunogen
Chemical/ Small Molecule corresponding to PIP2. Liposomes containing synthetic dipalmitoyl PtdIns(4,5)P2 or PtdIns(3,4,5)P3
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Positive control
- IHC-P: FFPE human kidney normal and FFPE mouse normal brain. ICC/IF: HepG2 cell line, Neuro2a cell line
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General notes
For testing in lipid dot blot assay, follow the protocol used in Thomas et al. Biochem Soc Trans 27:648-52 (1999) (PMID: 10917659, please see the References tab).
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
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 this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Some batches contain 6.97% L-Arginine as a stabilizing agent. For lot-specific buffer information, please contact our Scientific Support team. -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
2C11 -
Myeloma
Sp2/0-Ag14 -
Isotype
IgM -
Research areas
Associated products
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Compatible Secondaries
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab11039 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 |
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IHC-P |
Use a concentration of 1 - 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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ICC/IF | (2) |
Use a concentration of 1 - 5 µg/ml.
Fixation with 100% MeOH (5 min) or 4% PFA (10 min). |
ELISA |
Use at an assay dependent concentration.
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Neutralising |
Use at an assay dependent concentration.
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Notes |
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IHC-P
Use a concentration of 1 - 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
ICC/IF
Use a concentration of 1 - 5 µg/ml. Fixation with 100% MeOH (5 min) or 4% PFA (10 min). |
ELISA
Use at an assay dependent concentration. |
Neutralising
Use at an assay dependent concentration. |
Target
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Relevance
Phosphatidylinositol 4,5-biphosphate (PIP2) is a membrane phospholipid that has been implicated in a variety of cellular processes, including synaptic vesicle recycling and signal transduction pathways. PLCD4 hydrolyzes PIP2 to generate 2 second messenger molecules diacylglycerol (DAG) and inositol 1,4,5-trisphosphate (IP3).
Images
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Immunocytochemistry analysis of methanol-fixed 0.3% tritonX-100 permeabilized Bone Osteosarcoma Epithelial Cells staining with ab11039 at 1/200. Secondary antibody was Cy3® anti-mouse at 1/300 dilution. Samples were incubated with the primary antibody for 16 hours at 4°C. Blocking was done using 5% serum for 1 hour at 25°C.
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ICC/IF image of ab11039 stained human HepG2 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab11039, 5 µg/ml) overnight at +4°C.
The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgM (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue). This antibody also gave a positive IF result in HeLa, Hek293 and MCF7 cells.
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IHC image of PIP2 staining in mouse normal brain formalin fixed paraffin embedded tissue section. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins. The section was incubated with ab11039, 1µg/ml overnight at +4°C. An HRP-conjugated secondary (Ab98679, 1/1000 dilution) was used for 1hr at room temperature. The section was counterstained with haematoxylin and mounted with DPX.
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Immunofluorescence analysis of Human SaOS-2 (Human osteosarcoma) cells, staining PIP2 with ab11039. Cells were either unstimulated (upper panel) or stimulated with direct current (lower panel).
Cells were fixed in formaldehyde, permeabilized and then blocked with 1% BSA for 20 min. Cells were then incubated with a primary antibody (1/200) overnight at 4°C. A FITC-conjugated anti-mouse IgG was used as the secondary antibody. -
IHC image of PIP2 staining in a formalin fixed, paraffin embedded human normal kidney tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab11039, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (24)
ab11039 has been referenced in 24 publications.
- Lycas MD et al. Nanoscopic dopamine transporter distribution and conformation are inversely regulated by excitatory drive and D2 autoreceptor activity. Cell Rep 40:111431 (2022). PubMed: 36170827
- Gu J et al. CFTR Deficiency Affects Glucose Homeostasis via Regulating GLUT4 Plasma Membrane Transportation. Front Cell Dev Biol 9:630654 (2021). PubMed: 33659254
- Chen Y et al. Synaptotagmin-1 interacts with PI(4,5)P2 to initiate synaptic vesicle docking in hippocampal neurons. Cell Rep 34:108842 (2021). PubMed: 33730593
- Eaton-Fitch N et al. The effect of IL-2 stimulation and treatment of TRPM3 on channel co-localisation with PIP2 and NK cell function in myalgic encephalomyelitis/chronic fatigue syndrome patients. J Transl Med 19:306 (2021). PubMed: 34266470
- Petkovic M et al. TMEM16K is an interorganelle regulator of endosomal sorting. Nat Commun 11:3298 (2020). PubMed: 32620747