Skip to content

HE4 Recombinant Rabbit Monoclonal Antibody [JF62-09] - BSA and Azide free

Usd: 649

Specification

Bulk Order Quote | Customization Request

Catalog# HA750355

HE4 Recombinant Rabbit Monoclonal Antibody [JF62-09] - BSA and Azide free

Application

  • WB

  • IF-Cell

  • IF-Tissue

  • IHC-P

  • IP

  • FC

Reactivity

  • Human

  • Mouse

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

HE4 Recombinant Rabbit Monoclonal Antibody [JF62-09] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human HE4 aa 78-114 / 124.

Species Reactivity

Human, Mouse

Validated Applications

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

Molecular Weight

Predicted band size: 13 kDa

Positive Control

SW480 cell lysate, HepG2 cell lysate, OVCAR-3 cell lysate, mouse lung tissue lysate, mouse kidney tissue lysate, HepG2, mouse lung tissue.

Conjugation

unconjugated

Clone Number

JF62-09

Product Features

Form

Liquid

Concentration

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000

  • IF-Cell

  • 1:50-1:200

  • IF-Tissue

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:1,000

  • IP

  • Use at an assay dependent concentration.

Target

Function

HE4 (whey acidic protein (WAP)-type four-disulfide core-2, WFDC2) is a small secretory protein that may influence sperm maturation. HE4 gene expression is high in pulmonary epithelial cells and in some ovarian cancers. HE4 protein has a WAP motif that contains eight cysteines forming four disulfide bonds at the core of the protein. The WAP motif functions as a protease inhibitor in many of the family members that contain them.

Background References

1. Markovic-Lipkovski J et al. Variable Expression of Neural Cell Adhesion Molecule Isoforms in Renal Tissue: Possible Role in Incipient Renal Fibrosis. PLoS One 10:e0137028 (2015).

2. Zhuang H et al. Human epididymis protein 4 in association with Annexin II promotes invasion and metastasis of ovarian cancer cells. Mol Cancer 13:243 (2014).

Tissue Specificity

Expressed in a number of normal tissues, including male reproductive system, regions of the respiratory tract and nasopharynx. Highly expressed in a number of tumors cells lines, such ovarian, colon, breast, lung and renal cells lines. Initially described as being exclusively transcribed in the epididymis.

Subcellular Location

Secreted.

UNIPROT

SwissProt: Q14508 Human

SwissProt: Q9DAU7 Mouse

Synonyms

dJ461P17.6 antibody

EDDM4 antibody

epididymal protein 4 antibody

Epididymal secretory protein E4 antibody

Epididymis specific whey acidic protein type four disulfide core antibody

HE 4 antibody

Major epididymis specific protein E4 antibody

Major epididymis-specific protein E4 antibody

MGC57529 antibody

Putative protease inhibitor WAP5 antibody

Expand

Images

  • Western blot analysis of HE4 on different lysates with Rabbit anti-HE4 antibody (<a href="/products/HA750355" style="font-weight: bold;text-decoration: underline;">HA750355</a>) at 1/2,000 dilution.<br /><br />Lane 1: SW480 cell lysate (15 µg/Lane)<br />Lane 2: HepG2 cell lysate (15 µg/Lane)<br />Lane 3: OVCAR-3 cell lysate (15 µg/Lane)<br />Lane 4: Mouse lung tissue lysate (20 µg/Lane)<br />Lane 5: Mouse kidney tissue lysate (20 µg/Lane)<br /><br />Predicted band size: 13 kDa<br />Observed band size: 20 kDa<br /><br />Exposure time: 1 minute;<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/HA750355" style="font-weight: bold;text-decoration: underline;">HA750355</a>) at 1/2,000 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.

    Western blot analysis of HE4 on different lysates with Rabbit anti-HE4 antibody (HA750355) at 1/2,000 dilution.

    Lane 1: SW480 cell lysate (15 µg/Lane)
    Lane 2: HepG2 cell lysate (15 µg/Lane)
    Lane 3: OVCAR-3 cell lysate (15 µg/Lane)
    Lane 4: Mouse lung tissue lysate (20 µg/Lane)
    Lane 5: Mouse kidney tissue lysate (20 µg/Lane)

    Predicted band size: 13 kDa
    Observed band size: 20 kDa

    Exposure time: 1 minute;

    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 (HA750355) at 1/2,000 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.

  • Immunocytochemistry analysis of HepG2 cells labeling HE4 with Rabbit anti-HE4 antibody (<a href="/products/HA750355" style="font-weight: bold;text-decoration: underline;">HA750355</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-HE4 antibody (<a href="/products/HA750355" style="font-weight: bold;text-decoration: underline;">HA750355</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of HepG2 cells labeling HE4 with Rabbit anti-HE4 antibody (HA750355) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-HE4 antibody (HA750355) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Immunohistochemical analysis of paraffin-embedded mouse lung tissue using anti-HE4 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA750355" style="font-weight: bold;text-decoration: underline;">HA750355</a>, 1/50) for 30 minutes 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 lung tissue using anti-HE4 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750355, 1/50) for 30 minutes 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.

  • Flow cytometric analysis of HepG2 cells labeling HE4.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA750355" style="font-weight: bold;text-decoration: underline;">HA750355</a>, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of HepG2 cells labeling HE4.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA750355, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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