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Knockout Tested Rabbit Recombinant Monoclonal HE4 antibody. Suitable for IP, WB, IHC-P and reacts with Human samples. Cited in 15 publications.

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Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HE4 antibody [EPR16658] (AB200828), expandable thumbnail
  • Western blot - Anti-HE4 antibody [EPR16658] (AB200828), expandable thumbnail
  • Immunoprecipitation - Anti-HE4 antibody [EPR16658] (AB200828), expandable thumbnail
  • Western blot - Anti-HE4 antibody [EPR16658] (AB200828), expandable thumbnail
  • Western blot - Anti-HE4 antibody [EPR16658] (AB200828), expandable thumbnail

Publications

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPWBICC/IFIHC-P
Human
Tested
Tested
Not recommended
Tested

Tested
Tested

Species
Human
Dilution info
1/120
Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Tested
Tested

Species
Human
Dilution info
1/1000
Notes

-

Not recommended
Not recommended

Species
Human
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/1000
Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Target data

Function

Broad range protease inhibitor.

Alternative names

HE4, WAP5, WFDC2, WAP four-disulfide core domain protein 2, Epididymal secretory protein E4, Major epididymis-specific protein E4, Putative protease inhibitor WAP5

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Knockout Tested Rabbit Recombinant Monoclonal HE4 antibody. Suitable for IP, WB, IHC-P and reacts with Human samples. Cited in 15 publications.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
EPR16658
Purification technique
Affinity purification Protein A
Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

Human Epididymis Protein 4 (HE4) also known as WAP four-disulfide core domain protein 2 (WFDC2) is a small glycoprotein of approximately 13 kDa. HE4 is most highly expressed in the epithelial cells of reproductive tissues like the epididymis but also in tissues such as the respiratory and urogenital tracts. HE4 is secreted into the extracellular space where it functions and is found in normal range in different body fluids. This secreted protein has gained attention due to its use in various diagnostic tests such as the HE4 test and HE4 assay utilizing HE4 antibodies.

Biological function summary

HE4 participates in innate immune response and epithelial cell protection. HE4 protein plays a role in the modulation of antiprotease activity as it is involved in inhibiting serine proteases. It is not part of a larger protein complex but interacts with multiple protease signaling pathways. HE4 is often studied for its role in modulating protein activity at epithelial barriers contributing to tissue homeostasis and protective mechanisms against infections.

Pathways

HE4 interacts within signaling cascades such as the innate immune pathways and extracellular matrix organization. It links to proteins within these pathways like serine proteases and other WFDC family proteins. HE4 influences protease activity regulation an essential component of tissue remodeling and inflammatory response impacting overall pathway dynamics.

Associated diseases and disorders

HE4 has strong associations with ovarian cancer and endometrial cancer. Its overexpression in these malignancies makes it a valuable biomarker for disease diagnosis and prognosis. HE4 interacts with cancer-related proteins such as CA125 enhancing its utility in combinatory diagnostic approaches. The HE4 test and HE4 ELISA kit price underline its significance in clinical settings for monitoring these cancers where timely detection influences treatment outcomes.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

5 product images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HE4 antibody [EPR16658] (ab200828), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HE4 antibody [EPR16658] (ab200828)

    Immunohistochemical analysis of paraffin-embedded Human ovarian carcinoma labeling HE4 with ab200828 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.

    Cytoplasmic staining on Human ovarian carcinoma tissue is observed.

    Counter stained with Hematoxylin.
    Negative control: Used PBS instead of primary ab, secondary ab is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Western blot - Anti-HE4 antibody [EPR16658] (ab200828), expandable thumbnail

    Western blot - Anti-HE4 antibody [EPR16658] (ab200828)

    False colour image of Western blot: Anti-HE4 antibody [EPR16658] staining at 1/2000 dilution, shown in black; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab200828 was shown to bind specifically to HE4. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times before development with Optiblot (ECL reagent Anti-PKC delta (phospho S299) antibody [EPNCI119] ab133456) and imaged with 2 minutes 30 seconds exposure time. Secondary antibodies used were HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.

    All lanes: Western blot - Anti-HE4 antibody [EPR16658] (ab200828) at 1/2000 dilution

    Lane 1: OVCAR-3 cell lysate at 20 µg

    Lane 2: K562 cell lysate at 20 µg

    Performed under reducing conditions.

    Predicted band size: 13 kDa

    Observed band size: 20 kDa

  • Immunoprecipitation - Anti-HE4 antibody [EPR16658] (ab200828), expandable thumbnail

    Immunoprecipitation - Anti-HE4 antibody [EPR16658] (ab200828)

    HE4 was immunoprecipitated from 1mg of NIH:OVCAR-3 (Human ovary adenocarcinoma ) whole cell lysate with ab200828 at 1/120 dilution.

    Western blot was performed from the immunoprecipitate using ab200828 at 1/1000 dilution.

    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

    Lane 1: NIH:OVCAR-3 (Human ovary adenocarcinoma) whole cell lysate 10μg.

    Lane 2: NIH:OVCAR-3 (Human ovary adenocarcinoma) whole cell lysate following IP.

    Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab200828 in NIH:OVCAR-3 (human ovary adenocarcinoma) whole cell lysate.
    Blocking and dilution buffer and concentration: 5% NFDM/TBST. 10 second exposure.

    All lanes: Immunoprecipitation - Anti-HE4 antibody [EPR16658] (ab200828)

    Predicted band size: 13 kDa

    Observed band size: 17-25 kDa

  • Western blot - Anti-HE4 antibody [EPR16658] (ab200828), expandable thumbnail

    Western blot - Anti-HE4 antibody [EPR16658] (ab200828)

    Blocking/dilution buffer: 5% NFDM/TBST.

    The expression profile observed is consistent with what has been described in the literature PMID:15781627

    All lanes: Western blot - Anti-HE4 antibody [EPR16658] (ab200828) at 1/1000 dilution

    Lane 1: Human ovary cancer whole cell lysate at 10 µg

    Lane 2: NIH:OVCAR-3 (Human ovary adenocarcinoma) whole cell lysate at 10 µg

    Secondary

    All lanes: Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 13 kDa

    Observed band size: 17-25 kDa

    Exposure time: 1min

  • Western blot - Anti-HE4 antibody [EPR16658] (ab200828), expandable thumbnail

    Western blot - Anti-HE4 antibody [EPR16658] (ab200828)

    False colour image of Western blot: Anti-HE4 antibody [EPR16658] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab200828 was shown to bind specifically to HE4. A band was observed at 20 kDa in treated wild-type OVCAR-3 cell lysates with no signal observed at this size in WFDC2 knockout cell line ab275410 (knockout cell lysate ab275536). To generate this image, wild-type and WFDC2 knockout OVCAR-3 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

    All lanes: Western blot - Anti-HE4 antibody [EPR16658] (ab200828) at 1/1000 dilution

    Lane 1: Wild-type OVCAR-3 Treated Brefeldin A (Brefeldin A Solution (1,000X) ab193369) (5 ug/mL, 4 h), Monensin (Monensin Solution (1,000X) ab193381) (2 uM, 4 h) cell lysate at 20 µg

    Lane 2: Wild-type OVCAR-3 Vehicle Control Brefeldin A (Brefeldin A Solution (1,000X) ab193369) (0 ug/mL, 4 h), Monensin (Monensin Solution (1,000X) ab193381) (0 uM, 4 h) cell lysate at 20 µg

    Lane 3: WFDC2 knockout OVCAR-3 Treated Brefeldin A (Brefeldin A Solution (1,000X) ab193369) (5 ug/mL, 4 h), Monensin (Monensin Solution (1,000X) ab193381) (2 uM, 4 h) cell lysate at 20 µg

    Lane 4: WFDC2 knockout OVCAR-3 Vehicle Control Brefeldin A (Brefeldin A Solution (1,000X) ab193369) (0 ug/mL, 4 h), Monensin (Monensin Solution (1,000X) ab193381) (0 uM, 4 h) cell lysate at 20 µg

    Performed under reducing conditions.

    Observed band size: 20 kDa

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Product protocols

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