概述

  • 产品名称Anti-PIP2抗体[2C11]
    参阅全部 PIP2 一抗
  • 描述
    小鼠单克隆抗体[2C11] to PIP2
  • 特异性Reacts with PtdIns(4,5)P2, Ptd(4)P and Ptd(3,4,5)P3. To test antibody use a lipid blot assay as described in Thomas et al or a nuclear staining assay as in Osborne et al (full references under tab "References" at top of datasheet).
  • 经测试应用适用于: ICC/IF, ELISA, Neutralising, IPmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    Liposomes (prepared in PBS with lipid A, phosphatidylcholine (PC) and cholesterol) containing synthetic di-palmitoyl PtdIns(4,5)P2.

性能

应用

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.

应用 Ab评论 说明
ICC/IF 1/50. PubMed: 19124467
ELISA Use at an assay dependent concentration.
Neutralising Use at an assay dependent concentration.
IP Use at an assay dependent concentration.

靶标

  • 相关性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).

Anti-PIP2 antibody [2C11] 图像

  • 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.

  • 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.

Anti-PIP2 antibody [2C11] (ab11039)参考文献

This product has been referenced in:

See all 9 Publications for this product

Product Wall

We do share your concerns regarding mouse on mouse staining of kidney tissues. At Abcam we recommend following the attached mouse on mouse protocol tips to reduce background from this type of staining. While we do not have data regarding kidney tiss...

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Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HEK293)
Loading amount 20 µg
Specification HEK293
Gel Running Conditions Non-reduced Denaturing (10% gel)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
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Verified customer

提交于 Feb 01 2013

Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

The details you have kindly provided will enable us to investigate this ...

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Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from these antibodies.

I would like to reassure you that this antibodies are tested and covered by our 6 month guarantee for...

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Thank you for contacting us.

I personally don't have experience with using a freeze-thaw permeabilization, but from doing a quick literature search it does seem to be a good method for staining intracellular membrane proteins.

http...

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Thank you very much for your reply.
Since PIP2 seems to be ubiquitously expressed, I would expect most, if not all, cells to be positive controls. For ab11039, HepG2 may be the best positive control since the antibody has been tested in this cell ...

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Thank you again for your reply and for your patience.
I have followed-up with the lab and the secondary antibody mentioned in the image caption was actually an anti-mouse IgM, not IgG. We have corrected this in the caption. Thank you very much for...

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Thank you very much for your reply.
I'll need to check with the lab to confirm the type of secondary antibody that was used in this ICC/IF protocol. The anti-mouse IgG secondary antibody described in the image caption would still probably cross-re...

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Thank you very much for contacting us with your question.
I've looked through some literature, and it looks like PIP2 can be found in the nucleus as well as on the plasma membrane-
http://jcs.biologists.org/content/114/13/2501.long
http:...

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I am sorry that this antibody did not perform as stated on the datasheet. If payment has already been made on the ...

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