Anti-Phosphatidic acid phosphatase type 2B antibody [ZV002] (ab52581)

Overview

  • Product name
    Anti-Phosphatidic acid phosphatase type 2B antibody [ZV002]
    See all Phosphatidic acid phosphatase type 2B primary antibodies
  • Description
    Mouse monoclonal [ZV002] to Phosphatidic acid phosphatase type 2B
  • Host species
    Mouse
  • Specificity
    This antibody is specific for the ~33-35 kDa human VCIP (vascular endothelial growth factor and type I collagen inducible protein, phosphatidic acid phosphatase 2b, PAP2b) protein.
  • Tested applications
    Suitable for: WB, ELISAmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Recombinant protein derived from an internal region of human PPAP2b.

  • Positive control
    • Human PPAP2b-transfected 293 cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab52581 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 Use a concentration of 1 - 5 µg/ml. Predicted molecular weight: 35 kDa.
ELISA Use a concentration of 0.1 - 1 µg/ml.

Target

  • Function
    Catalyzes the conversion of phosphatidic acid (PA) to diacylglycerol (DG). In addition it hydrolyzes lysophosphatidic acid (LPA), ceramide-1-phosphate (C-1-P) and sphingosine-1-phosphate (S-1-P). The relative catalytic efficiency is LPA = PA > C-1-P > S-1-P. May be involved in cell adhesion and in cell-cell interactions.
  • Tissue specificity
    Ubiquitously expressed. Highly expressed in heart and placenta.
  • Sequence similarities
    Belongs to the PA-phosphatase related phosphoesterase family.
  • Post-translational
    modifications
    N-glycosylated. Contains high-mannose oligosaccharides.
  • Cellular localization
    Golgi apparatus > trans-Golgi network membrane. Cell membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • Dri 42 antibody
    • Dri42 antibody
    • Lipid phosphate phosphohydrolase 3 antibody
    • LPP3 antibody
    • LPP3_HUMAN antibody
    • PAP 2b antibody
    • PAP-2b antibody
    • PAP2 beta antibody
    • PAP2-beta antibody
    • PAP2b antibody
    • Phosphatidate phosphohydrolase type 2b antibody
    • Phosphatidic acid phosphatase 2b antibody
    • Ppap2b antibody
    • type 2 phosphatidic acid phosphatase beta antibody
    • vascular endothelial growth factor and type I collagen inducible antibody
    • Vascular endothelial growth factor and type I collagen inducible protein antibody
    • Vascular endothelial growth factor and type I collagen-inducible protein antibody
    • VCIP antibody
    see all

References

ab52581 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Question

BATCH NUMBER ZV002 ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM Multiple bands, all along the gel and where the expected band is supposed to be, we don't see it We are buying many antibodies and usually they work well, but this one doesn't work at all SAMPLE Total Membranes of mouse J774 cells PRIMARY ANTIBODY anti-PAP2B, Abcam cat#: ab52581, Mouse Monoclonal, clone ZV002, Dilution: 1/250 in either blocking agents Incubation: overnight at 4C for each conditions Wash: TBS or PBS 3 X 10 min. RT DETECTION METHOD Chemiluminescence Kit from Perkin-Elmer Different times of exposition POSITIVE AND NEGATIVE CONTROLS USED We didn't do any ANTIBODY STORAGE CONDITIONS antibody received on March 11th, 2008. Bought through Cedar Lane. Kept at -20C, until the day of experiment SAMPLE PREPARATION wash cell in PBS. Lysis Buffer: 8,55% sucrose solution containing 3mM Imidazole and Protease Inhibitors ( Complete 1X, from Roche ) Ultracentrifugation on 62% sucrose cushion and we have a total membrane fraction. AMOUNT OF PROTEIN LOADED we tried with 5ug, 10 ug, 15 ug and 20ug ELECTROPHORESIS/GEL CONDITIONS NuPage Bis-Tris 4-12% in reducing conditions Running Buffer: Tris glycins SDS TRANSFER AND BLOCKING CONDITIONS Nitrocellulose membrane, transfer time: 90 min, in Tris-glycine-SDS-Methanol buffer Tried 2 different blocking agents, for each primary antibody incubation, for each concentration of proteins: 1) 5% Milk-0,2%tween in either PBS or TBS 2) 5%BSA-0,2%Tween in either PBS or TBS SECONDARY ANTIBODY anti-mouse-HRP, from Jackson Immunochemicals Dilution: 1/3000 in either blocking agents Incubation: 30 min. RT Wash: Blocking agent 1 X 10 min. RT and TBS or PBS 3 X 10 min. at RT HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3X HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? the washing and blocking buffers have been changed, concentrations of proteins loaded on the gel have been changed

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Answer

I have just received a reply from the originator of the antibody, I hope this will also help you: "VCIP is also called PPAP2B: phosphatidic acid phosphatase type 2B. The protein encoded by this gene is a member of the phosphatidic acid phosphatase (PAP) family. PAPs convert phosphatidic acid to diacylglycerol, and function in de novo synthesis of glycerolipids as well as in receptor-activated signal transduction mediated by phospholipase D. This protein is a membrane glycoprotein localized at the cell plasma membrane. It has been shown to actively hydrolyze extracellular lysophosphatidic acid and short-chain phosphatidic acid. The expression of this gene is found to be enhanced by epidermal growth factor in Hela cells. Alternatively spliced transcript variants encoding the same protein have been described. The molecule is expressed highly in fetal pancreas. The molecule is about 33-35 kDa and mainly located in ER. 2 things that popped out at me were: That the gene is enhanced by EGF in HeLa cells and that the molecule is highly expressed in fetal pancreas. These facts and an internet search of research papers on VCIP may help your customer in his/her search for a positive control. About the customer's complaint of lots of background on their blots, are you working with them? They could try blocking overnight and reduce the % of BSA/milk in the antibody dilution buffer from 5% down to 3%-1%. Their protein of interest could be low expressing. Since they are not seeing the band of interest (at ~33-35 kDa) they might try loading ] 20 ug per lane. (We usually recommend as a starting concentration 10-30 ug/well.) They can go up to 100 ug, if they like. I also would try higher and lower primary ab concentration (from 1:100 to 1:500) and only incubate for 1-2 hr at RT (not overnight)."

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Question

BATCH NUMBER ZV002 ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM Multiple bands, all along the gel and where the expected band is supposed to be, we don't see it We are buying many antibodies and usually they work well, but this one doesn't work at all SAMPLE Total Membranes of mouse J774 cells PRIMARY ANTIBODY anti-PAP2B, Abcam cat#: ab52581, Mouse Monoclonal, clone ZV002, Dilution: 1/250 in either blocking agents Incubation: overnight at 4C for each conditions Wash: TBS or PBS 3 X 10 min. RT DETECTION METHOD Chemiluminescence Kit from Perkin-Elmer Different times of exposition POSITIVE AND NEGATIVE CONTROLS USED We didn't do any ANTIBODY STORAGE CONDITIONS antibody received on March 11th, 2008. Bought through Cedar Lane. Kept at -20C, until the day of experiment SAMPLE PREPARATION wash cell in PBS. Lysis Buffer: 8,55% sucrose solution containing 3mM Imidazole and Protease Inhibitors ( Complete 1X, from Roche ) Ultracentrifugation on 62% sucrose cushion and we have a total membrane fraction. AMOUNT OF PROTEIN LOADED we tried with 5ug, 10 ug, 15 ug and 20ug ELECTROPHORESIS/GEL CONDITIONS NuPage Bis-Tris 4-12% in reducing conditions Running Buffer: Tris glycins SDS TRANSFER AND BLOCKING CONDITIONS Nitrocellulose membrane, transfer time: 90 min, in Tris-glycine-SDS-Methanol buffer Tried 2 different blocking agents, for each primary antibody incubation, for each concentration of proteins: 1) 5% Milk-0,2%tween in either PBS or TBS 2) 5%BSA-0,2%Tween in either PBS or TBS SECONDARY ANTIBODY anti-mouse-HRP, from another company Dilution: 1/3000 in either blocking agents Incubation: 30 min. RT Wash: Blocking agent 1 X 10 min. RT and TBS or PBS 3 X 10 min. at RT HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3X HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? the washing and blocking buffers have been changed, concentrations of proteins loaded on the gel have been changed

Read More
Answer

Thank you for contacting us for technical support with ab52581. This antibody has so far not been tested in mouse samples though due to good homology of the immunogen with mouse PAP2B we would predict the antibody to cross react with the mouse protein. I would recommend to run a positive control of human PAP2B expressing cells. The problem of high background you are experiencing could be due to the dilution of primary antibody; I would like to suggest to dilute the antibody much more, and to dilute the antibody in TBST only (blocking only 1 hour)

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