Product datasheet Anti-EBAG9 Antibody Catalog Number: PB0584

BOSTER BIOLOGICAL TECHNOLOGY Special NO.1, International Enterprise Center, 2nd Guanshan Road, Wuhan, China

Web: www.boster.com.cn Phone: +86 27 67845390 Fax: +86 27 67845390 Email: [email protected]

Basic Information Product Name Anti-EBAG9 Antibody Name EBAG9 Source Rabbit IgG Species Reactivity human,mouse,rat Tested Application WB,IHC-P Contents 500ug/ml antibody with PBS ，0.02% NaN3 , 1mg BSA and 50% glycerol. Immunogen E.coli-derived human EBAG9 recombinant (Position: R31-S213). Human EBAG9 shares 97.8% and 94.5% amino acid (aa) sequence identity with mouse and rat EBAG9, respectively. Purification Immunogen affinity purified. Observed MW 34KD Dilution Ratios Western blot: 1:500-2000 Immunohistochemistry(Paraffin-embedded Section): 1:50-400 (Boiling the paraffin sections in 10mM citrate buffer,pH6.0,or PH8.0 EDTA repair liquid for 20 mins is required for the staining of formalin/paraffin sections.) Optimal working dilutions must be determined by end user.

Storage 12 months from date of receipt，-20℃ as supplied.6 months 2 to 8℃ after reconstitution. Avoid repeated freezing and thawing

Background Information Receptor-binding antigen expressed on SiSo cells is a protein that in humans is encoded by the EBAG9 gene. This gene was identified as an -responsive gene. Regulation of by estrogen is mediated by , which binds to the estrogen- responsive element found in the 5'-flanking region of this gene. And the encoded protein is a tumor-associated antigen that is expressed at high frequency in a variety of . Alternate splicing results in multiple transcript variants. A pseudogene of this gene has been defined on 10.

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FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC AND CLINICAL USE. 1 Product datasheet Anti-EBAG9 Antibody Catalog Number: PB0584

BOSTER BIOLOGICAL TECHNOLOGY Special NO.1, International Enterprise Center, 2nd Guanshan Road, Wuhan, China

Web: www.boster.com.cn Phone: +86 27 67845390 Fax: +86 27 67845390 Email: [email protected]

Selected Validation Data

Figure 1. Western blot Figure 2. IHC analysis of analysis of EBAG9 using EBAG9 using anti-EBAG9 anti-EBAG9 antibody antibody (PB0584).EBAG9 (PB0584). Electrophoresis was detected in paraffin- was performed on a embedded section of 5-20% SDS-PAGE gel at Mouse Testis Tissue. Heat 70V (Stacking gel) / 90V mediated antigen (Resolving gel) for 2-3 retrieval was performed hours. The sample well of in citrate buffer (pH6, each lane was loaded epitope retrieval solution) with 50ug of sample for 20 mins. The tissue under reducing section was blocked with conditions. Lane 1: Rat 10% goat serum. The Testis Tissue LysateLane tissue section was then 2: 22RV1 Whole Cell incubated with 1μg/ml LysateLane 3: HELA rabbit anti-EBAG9 Whole Cell LysateLane 4: Antibody (PB0584) MCF-7 Whole Cell overnight at 4°C. LysateLane 5: JURKAT Biotinylated goat anti- Whole Cell Lysate After rabbit IgG was used as Electrophoresis, secondary antibody and were transferred to a incubated for 30 minutes Nitrocellulose membrane at 37°C. The tissue at 150mA for 50-90 section was developed minutes. Blocked the using Strepavidin-Biotin- membrane with 5% Non- Complex (SABC)(Catalog fat Milk/ TBS for 1.5 hour # SA1022) with DAB as at RT. The membrane was the chromogen. incubated with rabbit anti- EBAG9 antigen affinity purified polyclonal antibody (Catalog # PB0584) at 0.5 μg/mL overnight at 4°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 EBAG9 at

FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC AND CLINICAL USE. 2 Product datasheet Anti-EBAG9 Antibody Catalog Number: PB0584

BOSTER BIOLOGICAL TECHNOLOGY Special NO.1, International Enterprise Center, 2nd Guanshan Road, Wuhan, China

Web: www.boster.com.cn Phone: +86 27 67845390 Fax: +86 27 67845390 Email: [email protected]

approximately 34KD. The expected band size for EBAG9 is at 34KD.

Figure 3. IHC analysis of Figure 4. IHC analysis of EBAG9 using anti-EBAG9 EBAG9 using anti-EBAG9 antibody (PB0584).EBAG9 antibody (PB0584).EBAG9 was detected in paraffin- was detected in paraffin- embedded section of Rat embedded section of Testis Tissue. Heat Human Prostatic Cancer mediated antigen Tissue. Heat mediated retrieval was performed antigen retrieval was in citrate buffer (pH6, performed in citrate epitope retrieval solution) buffer (pH6, epitope for 20 mins. The tissue retrieval solution) for 20 section was blocked with mins. The tissue section 10% goat serum. The was blocked with 10% tissue section was then goat serum. The tissue incubated with 1μg/ml section was then rabbit anti-EBAG9 incubated with 1μg/ml Antibody (PB0584) rabbit anti-EBAG9 overnight at 4°C. Antibody (PB0584) Biotinylated goat anti- overnight at 4°C. rabbit IgG was used as Biotinylated goat anti- secondary antibody and rabbit IgG was used as incubated for 30 minutes secondary antibody and at 37°C. The tissue incubated for 30 minutes section was developed at 37°C. The tissue using Strepavidin-Biotin- section was developed Complex (SABC)(Catalog using Strepavidin-Biotin- # SA1022) with DAB as Complex (SABC)(Catalog the chromogen. # SA1022) with DAB as the chromogen.

FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC AND CLINICAL USE. 3

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