Anti-Neuropilin 1 antibody [EPR3113] (ab81321)

Overview

  • Product nameAnti-Neuropilin 1 antibody [EPR3113]
    See all Neuropilin 1 primary antibodies
  • Description
    Rabbit monoclonal [EPR3113] to Neuropilin 1
  • Tested applicationsSuitable for: WB, IHC-P, IP, Flow Cyt, IHC-Fr, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Monkey, Marmoset (common)
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Neuropilin 1 aa 900 to the C-terminus (intracellular).
    (Peptide available as ab189308)

  • Positive control
    • WB: HUVEC and HepG2 cell lysates, human placenta, kidney and heart, mouse heart and kidney and rat heart and kidney tissue lysates. IHC-P: Human liver tissue. ICC/IF: MCF7 and HUVEC cells. Flow Cyt: HepG2 and MCF7 cells.
  • General notes

    Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Associated products

Applications

Our Abpromise guarantee covers the use of ab81321 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 1/1000 - 1/2000. Predicted molecular weight: 103 kDa.Can be blocked with Neuropilin 1 peptide (ab189308).
IHC-P 1/100 - 1/400. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

IP 1/100.
Flow Cyt 1/50 - 1/70.

The epitope that the antibody recognizes is intracellular. Fixation and permeabilization are necessary.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-Fr Use at an assay dependent concentration.
ICC/IF 1/250.

Target

  • FunctionThe membrane-bound isoform 1 is a receptor involved in the development of the cardiovascular system, in angiogenesis, in the formation of certain neuronal circuits and in organogenesis outside the nervous system. It mediates the chemorepulsant activity of semaphorins. It binds to semaphorin 3A, The PLGF-2 isoform of PGF, The VEGF-165 isoform of VEGF and VEGF-B. Coexpression with KDR results in increased VEGF-165 binding to KDR as well as increased chemotaxis. It may regulate VEGF-induced angiogenesis.
    The soluble isoform 2 binds VEGF-165 and appears to inhibit its binding to cells. It may also induce apoptosis by sequestering VEGF-165. May bind as well various members of the semaphorin family. Its expression has an averse effect on blood vessel number and integrity.
  • Tissue specificityThe expression of isoforms 1 and 2 does not seem to overlap. Isoform 1 is expressed by the blood vessels of different tissues. In the developing embryo it is found predominantly in the nervous system. In adult tissues, it is highly expressed in heart and placenta; moderately in lung, liver, skeletal muscle, kidney and pancreas; and low in adult brain. Isoform 2 is found in liver hepatocytes, kidney distal and proximal tubules.
  • Sequence similaritiesBelongs to the neuropilin family.
    Contains 2 CUB domains.
    Contains 2 F5/8 type C domains.
    Contains 1 MAM domain.
  • Cellular localizationSecreted and Cell membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • A5 protein antibody
    • BDCA4 antibody
    • BLOOD DENDRITIC CELL ANTIGEN 4 antibody
    • CD304 antibody
    • Neuropilin-1 antibody
    • Neuropilin1 antibody
    • NP1 antibody
    • NPN1 antibody
    • NRP 1 antibody
    • NRP antibody
    • NRP1 antibody
    • NRP1_HUMAN antibody
    • transmembrane receptor antibody
    • Vascular endothelial cell growth factor 165 receptor antibody
    • VEGF165R antibody
    see all

Anti-Neuropilin 1 antibody [EPR3113] images

  • Unpurified ab81321 staining Neuropilin 1 in rat brain tissue sections by Immunohistochemistry (frozen sections). Tissue was fixed with paraformaldehyde and then blocked with 10% serum for 1 hour at 27°C followed by incubation with the primary antibody, undiluted, for 14 hours at 4°C. An undiluted Cy3® conjugated donkey anti-rabbit was used as the secondary antibody.

    See Abreview

  • Unpurified ab81321 staining Neuropilin 1 in the COS1 fibroblast-like cell line derived from monkey kidney tissue by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with Triton X-100 0.1% and blocked with 10% serum for 60 minutes at 24°C. Samples were incubated with primary antibody (1/200) for 16 hours at 4°C. An Alexa Fluor® 488-conjugated goat anti-rabbit monoclonal(1/500) was used as the secondary antibody.

    See Abreview

  • All lanes : Anti-Neuropilin 1 antibody [EPR3113] (ab81321) at 1/10000 dilution (purified)

    Lane 1 : Mouse heart tissue lysate
    Lane 2 : Rat heart tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution

    Predicted band size : 103 kDa
    Observed band size : 120 kDa (why is the actual band size different from the predicted?)

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Anti-Neuropilin 1 antibody [EPR3113] (ab81321) at 1/10000 dilution (purified) + Human heart tissue lysate at 20 µg

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution

    Predicted band size : 103 kDa
    Observed band size : 120 kDa (why is the actual band size different from the predicted?)

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Anti-Neuropilin 1 antibody [EPR3113] (ab81321) at 1/2000 dilution (purified) + Human placenta tissue lysate at 20 µg

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution

    Predicted band size : 103 kDa
    Observed band size : 120 kDa (why is the actual band size different from the predicted?)

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling Neuropilin 1 with purified ab81321 at 1/400. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Immunocytochemistry/Immunofluorescence analysis of HUVEC cells labelling Neuropilin 1 with purified ab81321 at 1/250. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500) were also used.

    Control 1: primary antibody (1/250) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

    Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500).

  • Flow Cytometry analysis of MCF7 cells labelling Neuropilin 1 with purified ab81321 at 1/70 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

  • Immunohistochemical analysis of frozen human kidney tissue sections labelling Neurophilin 1 with ab81321 at a concentration of 1/100 for 18 hours at 4°C. The antibody was then blocked with a serum free protein block for 1 hour at 21°C. The secondary antibody used was a donkey anti-rabbit antibody conjugated to an Alexa488® dye inclubated at 1/400.

    See Abreview

  • All lanes : Anti-Neuropilin 1 antibody [EPR3113] (ab81321) at 1/1000 dilution (unpurified)

    Lane 1 : Human placenta lysate
    Lane 2 : HUVEC cell lysate
    Lane 3 : HepG2 cell lysate
    Lane 4 : Mouse heart tissue lysate
    Lane 5 : Mouse kidney tissue lysate
    Lane 6 : Rat heart tissue lysate
    Lane 7 : Rat kidney tissue lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

    Predicted band size : 103 kDa
    Observed band size : 120 kDa (why is the actual band size different from the predicted?)
  • Unpurified ab81321 staining Neuropilin 1 in mouse brain tissue sections by Immunohistochemistry (IHC-P - formaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 10% serum for 1 hour at room temperature; antigen retrieval was by heat mediation in citrate buffer (pH 6). Samples were incubated with primary antibody (1/100 in PBS + 2% blocking serum) for 16 hours at 4°C. A biotin-conjugated goat anti-rabbit IgG polyclonal (1/250) was used as the secondary antibody.

    See Abreview

  • ICC/IF image of unpurified ab81321 stained MCF7 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 (unpurified ab81321, 5µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-rabbit IgG - H&L, pre-adsorbed (ab96899) used at a 1/250 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) at a concentration of 1.43µM.

  • Overlay histogram showing HepG2 cells stained with unpurified ab81321 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (unpurified ab81321, 1/50 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (0.5µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a significantly decreased signal in HepG2 cells fixed with methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

References for Anti-Neuropilin 1 antibody [EPR3113] (ab81321)

This product has been referenced in:
  • Huang D  et al. VEGF-B inhibits hyperglycemia- and Macugen-induced retinal apoptosis. Sci Rep 6:26059 (2016). WB ; Rat . Read more (PubMed: 27189805) »
  • Wang S  et al. Interleukin-10 deficiency impairs regulatory T cell-derived neuropilin-1 functions and promotes Th1 and Th17 immunity. Sci Rep 6:24249 (2016). IHC (PFA fixed) . Read more (PubMed: 27075020) »

See all 20 Publications for this product

Product Wall

Application Immunohistochemistry (Frozen sections)
Sample Mouse Tissue sections (Kidney)
Permeabilization No
Specification Kidney
Blocking step Dako Serum-Free Protein Block as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 21°C
Fixative Acetone:Methanol 1:1
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Verified customer

Submitted Aug 10 2015

Application Immunohistochemistry (Frozen sections)
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 2.5% · Temperature: 25°C
Sample Marmoset (common) Tissue sections (Brain)
Specification Brain
Permeabilization No
Fixative Paraformaldehyde
Username

煜欽 林

Verified customer

Submitted Mar 10 2014



We can recommend ab125938 (Rabbit IgG, monoclonal [SP137] - isotype control) as isotype control for rabbit monoclonal primary antibodies.

http://www.abcam.com/index.html?datasheet=125938 (or use the following: http://www.abcam.com/i...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Monkey Cell lysate - whole cell (cos-1)
Loading amount 30 µg
Specification cos-1
Gel Running Conditions Reduced Denaturing (12%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
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Submitted Apr 02 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (NIH3T3)
Specification NIH3T3
Fixative Paraformaldehyde
Permeabilization Yes - 0.1% Triton
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 24°C
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Verified customer

Submitted Mar 19 2013


Nous avons 3 abreviews pour IHC-P avec ab81321, un chez l’humain (26 October 2012) et deux chez la souris (28 March 2012, 22 June 2010.) Ces trois abreviews montrent des bons résultats en utilisant l’anticorps à une dilut...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Monkey Cell (cos-1)
Specification cos-1
Fixative Paraformaldehyde
Permeabilization Yes - 0.1% Triton
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
Username

Abcam user community

Verified customer

Submitted Feb 28 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HUVEC)
Loading amount 30 µg
Specification HUVEC
Gel Running Conditions Reduced Denaturing (12)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
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Submitted Feb 28 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Rat Tissue lysate - whole (lung)
Loading amount 20 µg
Specification lung
Gel Running Conditions Reduced Denaturing (12)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
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Submitted Feb 28 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - whole (lung)
Loading amount 20 µg
Specification lung
Gel Running Conditions Reduced Denaturing (12)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Username

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Submitted Feb 28 2013

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