Anti-Doppel/PRND Picoband™ Antibody
|Reactivity||Human, Mouse, Rat|
|Product Name||Anti-Doppel/PRND Picoband™ Antibody|
|Storage & Handling||At -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time. Avoid repeated freezing and thawing.|
|Description||Rabbit IgG polyclonal antibody for Doppel detection. Tested with WB, IHC-P in Human;Mouse;Rat.|
|Cite This Product||Anti-Doppel/PRND Picoband™ Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # A05457)|
|Contents/Buffer||Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.|
|Immunogen||A synthetic peptide corresponding to a sequence of human Doppel (ATQAANQGEFQKPDNKLHQQVLWRLVQELCSLKH).|
|Reactivity||Human, Mouse, Rat|
Assay Dilutions Overview
Western blot, 0.1-0.5μg/ml
Immunohistochemistry(Paraffin-embedded Section), 0.5-1μg/ml
Boster's Secondary Antibodies And IHC, WB Kits
The following reagents are used to generate the images below.Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit (SV0002-1) for IHC(P).
Images And Assay Conditions
Figure 2. IHC analysis of Doppel using anti-Doppel antibody (A05457).
Doppel was detected in paraffin-embedded section of mouse testis 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 1Î¼g/ml rabbit anti-Doppel Antibody (A05457) overnight at 4Â°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37Â°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Figure 3. IHC analysis of Doppel using anti-Doppel antibody (A05457).
Doppel was detected in paraffin-embedded section of human testis 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 1Î¼g/ml rabbit anti-Doppel Antibody (A05457) overnight at 4Â°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37Â°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Figure 1. Western blot analysis of Doppel using anti-Doppel antibody (A05457).
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 SHG-44 whole cell lysate,
Lane 2: rat testis tissue lysates,
Lane 3: rat kidney tissue lysates,
Lane 4: mouse testis tissue lysates,
Lane 5: mouse kidney tissue lysates,
Lane 6: mouse brain 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-Doppel antigen affinity purified polyclonal antibody (Catalog # A05457) at 0.5 Î¼g/mL overnight at 4Â°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 (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Doppel at approximately 23KD. The expected band size for Doppel is at 20KD.
Protein Target Info (Source: Uniprot.org)
|Protein Name||prion like protein doppel|
|Tissue Specificity||Expressed in testis, in Sertoli cells, ejaculated spermatozoa and in seminal fluid (at protein level).|
|Alternative Names||Prion-like protein doppel; PrPLP; Prion protein 2; PRND; DPL; UNQ1830/PRO3443|
|Subcellular Localization||Cell membrane.|
*if product is indicated to react with multiple species, protein info is based on the human gene.
|Protein Function||Required for normal acrosome reaction and for normal male fertility (By similarity). Can bind Cu(2+) (PubMed:15218028, PubMed:20411530).|
|Research Areas||Human, Mouse, Rat
*You can search these to find other products in these research areas.
|Background||Prion protein 2 (dublet), also known as PRND, or Doppel protein, is a protein which in humans is encoded by the PRND gene. It is mapped to 20p13. This gene is found on chromosome 20, approximately 20 kbp downstream of the gene encoding cellular prion protein, to which it is biochemically and structurally similar. The protein encoded by this gene is a membrane glycosylphosphatidylinositol-anchored glycoprotein that is found predominantly in testis. Mutations in this gene may lead to neurological disorders.|
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