Product datasheet Anti-AAMP Catalog Number: PB9123

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-AAMP Antibody Name AAMP Source Rabbit IgG Species Reactivity human Tested Application WB, IHC-P, ICC/IF, FCM Contents 500 ug/ml antibody with PBS ，0.02% NaN3 , 1 mg BSA and 50% glycerol. Immunogen E.coli-derived human AAMP recombinant (Position: E235-R434). Purification Immunogen affinity purified. Observed MW 47KD Dilution Ratios Western blot: 1:500-2000 Immunohistochemistry(Paraffin-embedded Section): 1:50-400 Immunocytochemistry/Immunofluorescence (ICC/IF): 1:50-400 Flow cytometry (FCM): 1-3 μg/1x106 cells (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 AAMP, also known as Angio-associated, migratory cell protein, is a protein which in humans is encoded by the AAMP gene. It is mapped to 2q35. The gene product of AAMP is an immunoglobulin-type protein, which is found to be expressed strongly in endothelial cells, cytotrophoblasts, and poorly differentiated colon cells found in lymphatics. It has been demonstrated that an AAMP peptide containing the putative heparan sulfate-binding domain binds to heparin and mediates heparin-sensitive cell adhesion. AAMP plays a role in angiogenesis and cell migration. In smooth muscle cell migration, it may act through the RhoA pathway.

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FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC AND CLINICAL USE. 1 Product datasheet Anti-AAMP Antibody Catalog Number: PB9123

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. Western blot analysis of AAMP using analysis of AAMP using anti-AAMP antibody anti-AAMP antibody (PB9123). Electrophoresis (PB9123). Electrophoresis was performed on a was performed on a 5-20% SDS-PAGE gel at 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V 70V (Stacking gel) / 90V (Resolving gel) for 2-3 (Resolving gel) for 2-3 hours.Lane 1: hours. The sample well of Recombinant Human each lane was loaded AAMP Protein 0.5ng. with 50ug of sample After Electrophoresis, under reducing were transferred conditions. Lane 1: A431 to a Nitrocellulose Whole Cell LysateLane 2: membrane at 150mA for HELA Whole Cell 50-90 minutes. Blocked LysateLane 3: HEPG2 the membrane with 5% Whole Cell LysateLane 4: Non-fat Milk/ TBS for 1.5 MCF-7 Whole Cell hour at RT. The LysateAfter membrane was incubated Electrophoresis, proteins with rabbit anti-AAMP were transferred to a antigen affinity purified Nitrocellulose membrane polyclonal antibody at 150mA for 50-90 (Catalog # PB9123) at 0.5 minutes. Blocked the μg/mL overnight at 4°C, membrane with 5% Non- then washed with fat Milk/ TBS for 1.5 hour TBS-0.1%Tween 3 times at RT. The membrane was with 5 minutes each and incubated with rabbit anti- probed with a goat anti- AAMP antigen affinity rabbit IgG-HRP purified polyclonal secondary antibody at a antibody (Catalog # dilution of 1:10000 for 1.5 PB9123) at 0.5 μg/mL hour at RT. The signal is overnight at 4°C, then developed using an washed with Enhanced TBS-0.1%Tween 3 times Chemiluminescent with 5 minutes each and detection (ECL) kit probed with a goat anti- (Catalog # EK1002) with rabbit IgG-HRP Tanon 5200 system. 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 AAMP at approximately 47KD. The

FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC AND CLINICAL USE. 2 Product datasheet Anti-AAMP Antibody Catalog Number: PB9123

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]

expected band size for AAMP is at 47KD.

Figure 3. IHC analysis of Figure AAMP using anti-AAMP 4.Immunofluorescence antibody (PB9123).AAMP assay analysis of U2OS was detected in paraffin- cell at a dilution of 1:200 embedded section of dylight 550 goat anti- human mammary cancer rabbit IgG(red) blue tissue. Heat mediated Pseudocolor blue=DAPI. 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 1μg/ml rabbit anti-AAMP Antibody (PB9123) overnight at 4°C. Biotinylated goat anti- rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin- Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

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

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