Recombinant

Recombinant Anti-PLVAP/PV-1 antibody [MECA-32] (ab27853)

Submit a review Q&A (3)References (8)
Immunocytochemistry - Anti-PLVAP/PV-1 antibody [MECA-32] (ab27853)
  • Immunohistochemistry (Frozen sections) - Anti-PLVAP/PV-1 antibody [MECA-32] (ab27853)
  • Immunohistochemistry (Frozen sections) - Anti-PLVAP/PV-1 antibody [MECA-32] (ab27853)
  • Flow Cytometry - Anti-PLVAP/PV-1 antibody [MECA-32] (ab27853)
  • Anti-PLVAP/PV-1 antibody [MECA-32] (ab27853)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rat monoclonal [MECA-32] to PLVAP/PV-1
  • Suitable for: Flow Cyt, ICC, IHC-Fr
  • Reacts with: Mouse

Overview

  • Product name

    Anti-PLVAP/PV-1 antibody [MECA-32]
    See all PLVAP/PV-1 primary antibodies

  • Description

    Rat monoclonal [MECA-32] to PLVAP/PV-1

  • Host species

    Rat

  • Tested applications

    Suitable for: Flow Cyt, ICC, IHC-Frmore details

  • Species reactivity

    Reacts with: Mouse

  • Immunogen

    Tissue, cells or virus corresponding to Mouse PLVAP/PV-1.

  • Positive control

    • ICC and Flow Cyt: bEND.3 cells; IHC-Fr: Mouse large intestine tissue.

  • General notes

    This product has switched from a hybridoma to recombinant production method on 14th December 2020.

    Previously labelled as PLVAP.

    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
    For more information see here.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab27853 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
Use 0.1µg for 106 cells.
ICC
Use a concentration of 1 µg/ml.
IHC-Fr
Use a concentration of 1 µg/ml.
Notes
Flow Cyt
Use 0.1µg for 106 cells.
ICC
Use a concentration of 1 µg/ml.
IHC-Fr
Use a concentration of 1 µg/ml.

Target

  • Function

    Involved in the formation of stomatal and fenestral diaphragms of caveolae. May function in microvascular permeability.

  • Tissue specificity

    Expressed in lung, kidney, heart, aorta, placenta, muscle, pituitary gland, adrenals, mammary gland, bladder, lymph node, bone marrow, trachea, digestive tract, liver and tumor-associated endothelium.

  • Cellular localization

    Cell membrane. Membrane > caveola. Cytoplasm > perinuclear region. Membrane-associated protein of caveolae. Found in fenestral and stomatal diaphragms in fenestrated endothelia and transendothelial channels. Also colocalized with CAV1 in perinuclear region.
  • Information by UniProt

  • Database links

    • Alternative names

      • FELS antibody
      • Fenestrated endothelial linked structure protein antibody
      • Fenestrated endothelial-linked structure protein antibody
      • gp68 antibody
      • Hypothetical protein DKFZp667O074 antibody
      • Meca 32 antigen antibody
      • MECA32 antigen antibody
      • Panendothelial cell antigen antibody
      • Plasmalemma vesicle associated protein antibody
      • Plasmalemma vesicle protein 1 antibody
      • Plasmalemma vesicle-associated protein antibody
      • Plvap antibody
      • PLVAP_HUMAN antibody
      • PV 1 antibody
      • PV-1 antibody
      • PV1 antibody
      • PV1 protein antibody
      see all

    Images

    • Immunocytochemistry - Anti-PLVAP/PV-1 antibody [MECA-32] (ab27853)
      Immunocytochemistry - Anti-PLVAP/PV-1 antibody [MECA-32] (ab27853)

      ab27853 staining PLVAP/PV-1 in b.End3 cells. The cells were fixed with 4% paraformaldehyde (10 minutes), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1 hour. The cells were then incubated overnight at 4°C with ab27853 at 2 µg/mL and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150165, Goat polyclonal Secondary Antibody to Rat IgG - H&L (Alexa Fluor® 488), pre-absorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue). Also suitable in cells fixed with 100% methanol (5 minutes).

      Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

    • Immunohistochemistry (Frozen sections) - Anti-PLVAP/PV-1 antibody [MECA-32] (ab27853)
      Immunohistochemistry (Frozen sections) - Anti-PLVAP/PV-1 antibody [MECA-32] (ab27853)

      IHC image of PLVAP/PV-1 staining in a section of frozen normal mouse large intestine performed on a Leica BOND™ system using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab27853, 1 µg/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. The inset secondary-only control image is taken from an identical assay without primary antibody.

      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.

    • Immunohistochemistry (Frozen sections) - Anti-PLVAP/PV-1 antibody [MECA-32] (ab27853)
      Immunohistochemistry (Frozen sections) - Anti-PLVAP/PV-1 antibody [MECA-32] (ab27853)

      IHC image of PLVAP/PV-1 staining in a section of frozen normal mouse brain performed on a Leica BOND™ system using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab27853, 1 µg/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. The inset secondary-only control image is taken from an identical assay without primary antibody. Mouse brain tissue is a negative control tissue, showing no staining of the primary antibody.

      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.

    • Flow Cytometry - Anti-PLVAP/PV-1 antibody [MECA-32] (ab27853)
      Flow Cytometry - Anti-PLVAP/PV-1 antibody [MECA-32] (ab27853)

      Flow cytometry overlay histogram showing bEND.3 (mouse brain endothelioma cell line) cells stained with ab27853 (red line). The cells were incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab27853) (1x106 in 100 µL at 1 µg/mL) for 30 min on ice.

      The secondary antibody Goat anti-rat IgG H&L (Alexa Fluor® 488, pre-adsorbed) (ab150165) was used at 1/2000 for 30 min on ice.

      Isotype control antibody (black line) was Rat IgG2aκ (ab18450) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

      Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter.

    • Anti-PLVAP/PV-1 antibody [MECA-32] (ab27853)
      Anti-PLVAP/PV-1 antibody [MECA-32] (ab27853)

    References (8)

    Publishing research using ab27853? Please let us know so that we can cite the reference in this datasheet.

    ab27853 has been referenced in 8 publications.

    • Tian M  et al. NL1 expression level in Nrx1ß and the excitability of PV interneurons in mice with POCD. Exp Ther Med 17:3117-3123 (2019). PubMed: 30936983
    • Englinger B  et al. Intrinsic fluorescence of the clinically approved multikinase inhibitor nintedanib reveals lysosomal sequestration as resistance mechanism in FGFR-driven lung cancer. J Exp Clin Cancer Res 36:122 (2017). PubMed: 28882160
    • Wang XL  et al. VEGFR-3 blocking deteriorates inflammation with impaired lymphatic function and different changes in lymphatic vessels in acute and chronic colitis. Am J Transl Res 8:827-41 (2016). IHC-Fr ; Mouse . PubMed: 27158372
    • Elgueta R  et al. Endothelial Plasmalemma Vesicle-Associated Protein Regulates the Homeostasis of Splenic Immature B Cells and B-1 B Cells. J Immunol 197:3970-3981 (2016). Flow Cyt . PubMed: 27742829
    • Menezes MJ  et al. The extracellular matrix protein laminin a2 regulates the maturation and function of the blood-brain barrier. J Neurosci 34:15260-80 (2014). PubMed: 25392494
    • Aguilera KY & Brekken RA Hypoxia Studies with Pimonidazole in vivo. Bio Protoc 4:N/A (2014). PubMed: 27453908
    • Orosz CG  et al. Treatment with anti-vascular cell adhesion molecule 1 monoclonal antibody induces long-term murine cardiac allograft acceptance. Transplantation 56:453-60 (1993). PubMed: 7689262
    • Pelletier RP  et al. Analysis of inflammatory endothelial changes, including VCAM-1 expression, in murine cardiac grafts. Transplantation 55:315-20 (1993). PubMed: 7679529

    Customer reviews and Q&As

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    1-3 of 3 Abreviews or Q&A

    Question

    ab2350: IHC-Fr with mouse lung and liver (20 and 90 um sections)
    liver: not intense but correct staining
    lung: no signal
    Ab: 1/50, 1/25 o/n 4 degree
    blocking: 55 serum
    secondary: Alexa488, works well
    perm step: TX100
    fix step: acetone 10 min
    send IHC-Fr protocol for ab27853

    ab53444: IHC-Fr with mouse lung and spleen
    high background
    Ab: 1/50
    suggested: use less antibodies and incubate shorter, block longer and more (10% serum)
    send IHC-Fr protocol

    Read More
    Answer

    Thank you for your patience.

    For ab27853, it is as I had thought in my last email.The application was added based on publications showing that [MECA-32] has worked in IHC-Fr, e.g.

    "The Adnectin CT-322 is a novel VEGF receptor 2 inhibitor that decreases tumor burden in an orthotopic mouse model of pancreatic cancer" by Dineen et al., BMC Cancer 2008, 8:352 (see attachment).

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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    Question

    ab2350: IHC-Fr with mouse lung and liver (20 and 90 um sections)
    liver: not intense but correct staining
    lung: no signal
    Ab: 1/50, 1/25 o/n 4 degree
    blocking: 55 serum
    secondary: Alexa488, works well
    perm step: TX100
    fix step: acetone 10 min
    send IHC-Fr protocol for ab27853

    ab53444: IHC-Fr with mouse lung and spleen
    high background
    Ab: 1/50
    suggested: use less antibodies and incubate shorter, block longer and more (10% serum)
    send IHC-Fr protocol

    Read More
    Answer

    Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

    I am sorry the product ab2350 did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number xxx with ab27853.

    To check the status of the order please contact our Customer Service team and reference this number.

    Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

    As for the IHC-Fr protocols:


    For ab27853, I will need to check with the supplying lab if they have any details on that. As far as I know, it was reported to them by other researchers that the antibody works in IHC-Fr. I will let you know what I find out.

    I have attached our IHC-Fr protocols to help you further in the meantime.


    For ab53444,we have the following protocol on file:

    Indirect Immunostaining of Frozen Tissue Sections

    For use with unconjugated monoclonal and polyclonal antibodies.
    Please note that a certain level of technical skill and immunological knowledge is required for the successful design and implementation of these techniques - these are guidelines only and may need to be adjusted for particular applications.
    Reagents
    1. 0.3% H2O2‎, 70% methanol in TBS
    1 ml of 30% H2O2‎ per 100 ml methanol/TBS.
    2. TBS stock solution (10x)
    NaCl, 87.66 g
    Tris base, 60.55 g
    Distilled water, 1 liter
    Adjust pH to 7.4 using concentrated HCl.
    Method
    In most cases, we recommend that tissues are snap-frozen in liquid nitrogen, then prepared as 4-6 μm sections using a cryostat.
    Allow sections to air dry for at least 1 hour.
    Fix sections in dry acetone for 15 minutes. Allow to evaporate for 10 minutes.
    Block endogenous peroxidase (if necessary) by immersing slides in 0.3% H2O2‎ in 70% methanol/TBS for 30 minutes. Wash once in TBS.
    Incubate sections for 10 minutes in 10% normal serum from the species in which the secondary was raised. Tap excess serum off the slides before staining.
    Incubate sections with primary antibody for at least 30 minutes at room temperature in a humid chamber, or overnight at 4°C. Wash 3 times in TBS.
    Add enzyme-conjugated secondary antibody at the recommended dilution (see specific datasheet for details). Incubate for at least 30 minutes at room temperature. Wash 3 times in TBS.
    Incubate with the appropriate substrate solution for the recommended period of time (it is suggested to useDAB substrate with HRP-conjugated antibodies, and Fast Red/Napthol AS-MX for Alkaline Phosphatase-conjugated antibodies). Wash once in water.
    Counterstain with hematoxylin for 1-10 minutes. “Blue” with running water for 5 minutes. Then wash.
    Mount in aqueous mounting medium, or alternatively dehydrate through a graded series of alcohols and xylene/solvent, and mount in synthetic mountant.

    Notes
    - Do not allow slides to dry out after the fixation step, as drying will result in damage to the tissue structure.
    - Beware, certain substrates are soluble in alcohol – please refer to manufacturer's information for details.
    - Appropriate controls should always be carried out. It may be useful to include a sample in which no primary antibody is used at all, in order to determine any non-specific binding of the secondary reagent to the target tissue.

    In addition, you can also check the Abreviews section where 7 IHC-Fr reviews had been submitted by other customers:
    https://www.abcam.com/CD68-antibody-FA-11-ab53444-reviews.html




    I wish you the best of luck with your research.

    Please let me know if there is anything else I can do for you.

    Read More

    Question

    I would like to order Anti ACVRL antibody #37807 from abcam for staining tumour samples. Since ALK is a endothelial specific marker...I should see only blood vessels positive, however, in the data sheet of your product you have photographs showing all the cells positive in a carcinoma. Do u have any references saying that ALK-1 is present on all or nearly all cells of the tumour.

    Read More
    Answer

    Thank you for contacting us. While I am unaware of any references describingACVRL1 expression in tumors, the human protein atlas (linked below) does show that "distinct cytoplasmic staining with a granular pattern was observed in thyroid cancers, malignant lymphomas and in a few malignant melanomas, colorectal, gastric, pancreatic and renal cancers. Malignant cells in general were negative." http://www.proteinatlas.org/ENSG00000139567/cancer Additionally, because ACVRL1 is88%homologous betweenmouse and human, it will be difficult to find an antibody to that target which will react with mouse and not human epithelial cells. If you are looking to specifically detect mouse endothelial cells, you may be interested in the PLVAP antibody clone MECA-32 (ab27853), which is described in http://www.ncbi.nlm.nih.gov/pubmed/7626790?dopt=AbstractPlus. I hope this helps, please let me know if you need any additional information.

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