Western Blot (WB)
(Figure 1. Western blot analysis of HE4 using anti-HE4 antibody.Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel)/90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: human Hela whole cell lysates,Lane 2: human MDA-MB-231 whole cell lysates,Lane 3: human MDA-MB-453 whole cell lysates,Lane 4: rat thymus tissue lysates,Lane 5: mouse testis tissue lysates,Lane 6: mouse thymus tissue lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-HE4 antigen affinity purified polyclonal antibody at 0.5 ug/mL overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for HE4 at approximately 15KD. The expected band size for HE4 is at 13KD.)
Rabbit HE4/WFDC2 Polyclonal Antibody | anti-HE4 antibody
Anti-HE4/WFDC2 Antibody
IHC-P: 0.5-1 mug/ml
Direct ELISA: 0.1-0.5 mug/ml
Tested Species: In-house tested species with positive results. By Heat: Boiling the paraffin sections in 10mM citrate buffer, pH6.0, for 20mins is required for the staining of formalin/paraffin sections.
Avoid repeated freezing and thawing.
Western Blot (WB)
(Figure 1. Western blot analysis of HE4 using anti-HE4 antibody.Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel)/90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: human Hela whole cell lysates,Lane 2: human MDA-MB-231 whole cell lysates,Lane 3: human MDA-MB-453 whole cell lysates,Lane 4: rat thymus tissue lysates,Lane 5: mouse testis tissue lysates,Lane 6: mouse thymus tissue lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-HE4 antigen affinity purified polyclonal antibody at 0.5 ug/mL overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for HE4 at approximately 15KD. The expected band size for HE4 is at 13KD.)
Western Blot (WB)
(Figure 1. Western blot analysis of HE4 using anti-HE4 antibody.Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel)/90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: human Hela whole cell lysates,Lane 2: human MDA-MB-231 whole cell lysates,Lane 3: human MDA-MB-453 whole cell lysates,Lane 4: rat thymus tissue lysates,Lane 5: mouse testis tissue lysates,Lane 6: mouse thymus tissue lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-HE4 antigen affinity purified polyclonal antibody at 0.5 ug/mL overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for HE4 at approximately 15KD. The expected band size for HE4 is at 13KD.)
Immunohistochemistry (IHC)
(Figure 3. IHC analysis of HE4 using anti-HE4 antibody.HE4 was detected in paraffin-embedded section of mouse spleen tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/mL rabbit anti-HE4 antibody.HE4 was detected in paraffin-embedded section of mouse spleen tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/mL rabbit anti-HE4 antibody.HE4 was detected in paraffin-embedded section of rat spleen tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-HE4 Antibody overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.)
Immunohistochemistry (IHC)
(Figure 3. IHC analysis of HE4 using anti-HE4 antibody.HE4 was detected in paraffin-embedded section of mouse spleen tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/mL rabbit anti-HE4 antibody.HE4 was detected in paraffin-embedded section of mouse spleen tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/mL rabbit anti-HE4 antibody.HE4 was detected in paraffin-embedded section of rat spleen tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-HE4 Antibody overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.)
Background: WAP four-disulfide core domain protein 2, also known as Human Epididymis Protein 4 (HE4), is a protein that in humans is encoded by the WFDC2 gene. This gene encodes a protein that is a member of the WFDC domain family. The WFDC domain, or WAP Signature motif, contains eight cysteines forming four disulfide bonds at the core of the protein, and functions as a protease inhibitor in many family members. This gene is expressed in pulmonary epithelial cells, and was also found to be expressed in some ovarian cancers. The encoded protein is a small secretory protein, which may be involved in sperm maturation.
NCBI and Uniprot Product Information
NCBI Description
This gene encodes a protein that is a member of the WFDC domain family. The WFDC domain, or WAP Signature motif, contains eight cysteines forming four disulfide bonds at the core of the protein, and functions as a protease inhibitor in many family members. This gene is expressed in pulmonary epithelial cells, and was also found to be expressed in some ovarian cancers. The encoded protein is a small secretory protein, which may be involved in sperm maturation. [provided by RefSeq, Jul 2008]
Uniprot Description
Broad range protease inhibitor.
Research Articles on HE4
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Product Notes
The HE4 wfdc2 (Catalog #AAA1751430) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-HE4/WFDC2 Antibody reacts with Human, Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's HE4/WFDC2 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunohistochemistry (IHC) Paraffin, Direct ELISA (EIA). WB: 0.1-0.5 mug/ml IHC-P: 0.5-1 mug/ml Direct ELISA: 0.1-0.5 mug/ml Tested Species: In-house tested species with positive results. By Heat: Boiling the paraffin sections in 10mM citrate buffer, pH6.0, for 20mins is required for the staining of formalin/paraffin sections. Researchers should empirically determine the suitability of the HE4 wfdc2 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "HE4/WFDC2, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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