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Neuropilin 1 Recombinant Rabbit Monoclonal Antibody [ST05-30]

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Specification

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Catalog# ET1609-69

Neuropilin 1 Recombinant Rabbit Monoclonal Antibody [ST05-30]

Application

  • WB

  • IF-Cell

  • IF-Tissue

  • IHC-P

  • IP

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

Neuropilin 1 Recombinant Rabbit Monoclonal Antibody [ST05-30]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human Neuropilin-1 aa 881-923 / 923.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IF-Tissue, IHC-P, IP

Molecular Weight

Predicted band size: 103 kDa

Positive Control

U-87 MG cell lysate, A549 cell lysate, MDA-MB-231 cell lysate, mouse brain tissue lysate, rat brain tissue lysate, human liver tissue, mouse heart tissue lysate, rat heart tissue lysate, SHG-44, MCF-7, HUVEC, human kidney tissue, mouse kidney tissue.

Conjugation

unconjugated

Clone Number

ST05-30

RRID

AB_3069878

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000-1:5,000

  • IF-Cell

  • 1:50-1:200

  • IF-Tissue

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

  • IP

  • Use at an assay dependent concentration.

Target

Function

Neuropilin is a type I transmembrane receptor that has been implicated in aspects of axon growth and guidance and has been shown to act as a high affinity receptor for class III semaphorins and vascular endothelial growth factor (VEGF). A closely related protein, neuropilin-2, shares a common domain structure and signifcant homology with neuropilin and also acts as a receptor for the class III semaphorins and VEGF. Both neuropilins are involved in regulating many physiological pathways including axonal guidance and angiogenesis, however they exhibit differential expression in the adult vasculature. Neuropilin-2 is polysialylated and expressed on the surface of dendritic cells. It is also expressed by venous and lymphatic endothelium. Neuropilin is expressed predominantly by arterial endothelium.

Background References

1. Liu F et al. Prostate cancer cells induce osteoblastic differentiation via semaphorin 3A. Prostate 75:370-80 (2015).

2. Linthicum FH et al. The periductal channels of the endolymphatic duct, hydrodynamic implications. Otolaryngol Head Neck Surg 150:441-7 (2014).

Sequence Similarity

Belongs to the neuropilin family.

Tissue Specificity

The 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.

Subcellular Location

Cell membrane, Secreted.

UNIPROT

SwissProt: O14786 Human

SwissProt: P97333 Mouse

SwissProt: Q9QWJ9 Rat

Synonyms

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

Expand

Images

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of Neuropilin 1 on different lysates with Rabbit anti-Neuropilin 1 antibody (<a href="/products/ET1609-69" style="font-weight: bold;text-decoration: underline;">ET1609-69</a>) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution.<br /><br />Lane 1: U-87 MG cell lysate<br />Lane 2: A549 cell lysate<br />Lane 3: SK-Br-3 cell lysate (low expression)<br />Lane 4: MDA-MB-231 cell lysate<br />Lane 5: Mouse brain tissue lysate<br />Lane 6: Rat brain tissue lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 103 kDa<br />Observed band size: 130 kDa<br /><br />Exposure time: 35 seconds; ECL: K1802;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1609-69" style="font-weight: bold;text-decoration: underline;">ET1609-69</a>) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution were used in 5% BSA at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Relative expression (RE)

    Western blot analysis of Neuropilin 1 on different lysates with Rabbit anti-Neuropilin 1 antibody (ET1609-69) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution.

    Lane 1: U-87 MG cell lysate
    Lane 2: A549 cell lysate
    Lane 3: SK-Br-3 cell lysate (low expression)
    Lane 4: MDA-MB-231 cell lysate
    Lane 5: Mouse brain tissue lysate
    Lane 6: Rat brain tissue lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 103 kDa
    Observed band size: 130 kDa

    Exposure time: 35 seconds; ECL: K1802;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1609-69) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution were used in 5% BSA at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-Neuropilin 1 antibody (<a href="/products/ET1609-69" style="font-weight: bold;text-decoration: underline;">ET1609-69</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1609-69" style="font-weight: bold;text-decoration: underline;">ET1609-69</a>) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-Neuropilin 1 antibody (ET1609-69) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1609-69) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of Neuropilin 1 on different lysates with Rabbit anti-Neuropilin 1 antibody (<a href="/products/ET1609-69" style="font-weight: bold;text-decoration: underline;">ET1609-69</a>) at 1/5,000 dilution.<br /><br />Lane 1: A549 cell lysate<br />Lane 2: MDA-MB-231 cell lysate<br />Lane 3: SK-Br-3 cell lysate (low expression)<br />Lane 4: Mouse brain tissue lysate<br />Lane 5: Mouse heart tissue lysate<br />Lane 6: Rat brain tissue lysate<br />Lane 7: Rat heart tissue lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 103 kDa<br />Observed band size: 130 kDa<br /><br />Exposure time: Lane 1-3: 1 minute 21 seconds; Lane 4-7: 43 seconds;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1609-69" style="font-weight: bold;text-decoration: underline;">ET1609-69</a>) at 1/5,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:100,000 dilution was used for 1 hour at room temperature.

    ☑ Relative expression (RE)

    Western blot analysis of Neuropilin 1 on different lysates with Rabbit anti-Neuropilin 1 antibody (ET1609-69) at 1/5,000 dilution.

    Lane 1: A549 cell lysate
    Lane 2: MDA-MB-231 cell lysate
    Lane 3: SK-Br-3 cell lysate (low expression)
    Lane 4: Mouse brain tissue lysate
    Lane 5: Mouse heart tissue lysate
    Lane 6: Rat brain tissue lysate
    Lane 7: Rat heart tissue lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 103 kDa
    Observed band size: 130 kDa

    Exposure time: Lane 1-3: 1 minute 21 seconds; Lane 4-7: 43 seconds;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1609-69) at 1/5,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.

  • ICC staining of Neuropilin 1 in SHG-44 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (<a href="/products/ET1609-69" style="font-weight: bold;text-decoration: underline;">ET1609-69</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

    ICC staining of Neuropilin 1 in SHG-44 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1609-69, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

  • ICC staining of Neuropilin 1 in MCF-7 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (<a href="/products/ET1609-69" style="font-weight: bold;text-decoration: underline;">ET1609-69</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

    ICC staining of Neuropilin 1 in MCF-7 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1609-69, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

  • ICC staining of Neuropilin 1 in HUVEC cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (<a href="/products/ET1609-69" style="font-weight: bold;text-decoration: underline;">ET1609-69</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

    ICC staining of Neuropilin 1 in HUVEC cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1609-69, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

  • Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Neuropilin 1 antibody (<a href="/products/ET1609-69" style="font-weight: bold;text-decoration: underline;">ET1609-69</a>) at 1/50 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1609-69" style="font-weight: bold;text-decoration: underline;">ET1609-69</a>) at 1/50 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Neuropilin 1 antibody (ET1609-69) at 1/50 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1609-69) at 1/50 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-Neuropilin 1 antibody (<a href="/products/ET1609-69" style="font-weight: bold;text-decoration: underline;">ET1609-69</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1609-69" style="font-weight: bold;text-decoration: underline;">ET1609-69</a>) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-Neuropilin 1 antibody (ET1609-69) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1609-69) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of Neuropilin 1 on different lysates with Rabbit anti-Neuropilin 1 antibody (<a href="/products/ET1609-69" style="font-weight: bold;text-decoration: underline;">ET1609-69</a>) at 1/5,000 dilution.<br /><br />Lane 1: MDA-MB-231-si NT cell lysate<br />Lane 2: MDA-MB-231-si Neuropilin 1 cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 103 kDa<br />Observed band size: 130 kDa<br /><br />Exposure time: 1 minute 2 seconds;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1609-69" style="font-weight: bold;text-decoration: underline;">ET1609-69</a>) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of Neuropilin 1 on different lysates with Rabbit anti-Neuropilin 1 antibody (ET1609-69) at 1/5,000 dilution.

    Lane 1: MDA-MB-231-si NT cell lysate
    Lane 2: MDA-MB-231-si Neuropilin 1 cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 103 kDa
    Observed band size: 130 kDa

    Exposure time: 1 minute 2 seconds;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1609-69) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation

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Conjugate: unconjugated

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