Ab-Match Assembly Human PAP1 (REG3a) Kit

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1 CODE No. 323 For Research Use Only, Not for use in diagnostic procedures. Ab-Match Assembly Human PAP1 (REG3α) Kit For technical material or related information, please refer to Advantage By using Ab-Match Assembly Human PAP1 (REG3α) Kit in combination with the Ab-Match Universal Kit (MBL; code no. 31), human PAP1 can be measured quantitatively and specifically. The two kits are sufficient to produce one 96-well microplate. This product is intended for research use only. Materials provided 1 96 wells Name Materials Quantity Dilution ratio PAP1 standard Recombinant human PAP1* 2 µl 1 vial 1:1 1:32 or 1:64 Anti-PAP1 capture antibody Mouse anti-pap1 monoclonal antibody (IgG) (% Glycerol/PBS) 6 µl 1 vial 1:2 Anti-PAP1 detection antibody Biotin conjugated rabbit anti-pap1 polyclonal antibody 11 µl 1 vial 1:11 Streptavidin conj. peroxidase Streptavidin conjugated peroxidase (SA-HRP) 11 µl 1 vial 1:11 * The concentration is printed to the label. To avoid repeated freezing and thawing, prepare aliquots. Note: Kit components must be brought to room (2-3 C) prior to use. Protocol Coating step Add 1 µl diluted capture antibody to each well Incubate over night at 2-8 C Blocking step Wash 2 times Add 2 µl of Blocking Agent to each well Incubate 1 hour at room Sample Dump out the contents of the wells Add 1 µl of samples or standards to each well Incubate 1 hour at room Detection antibody Wash 4 times Add 1 µl of diluted detection antibody to each well Incubate 1 hour at room SA-HRP step Wash 4 times Add 1 µl of diluted SA-HRP to each well Incubate 3 minutes at room Wash 4 times Dump out the contents of the wells Add 1 µl of Solution to each well Incubate 3 minutes at room Reading Add 1 µl of Stop Solution to each well Read absorbance at 4 nm Storage and Stability All kit components must be stored at -2 ~ -3 C. All reagents are stable for 12 months after shipment. Related product 31 Ab-Match Universal Kit MEDICAL & BIOLOGICAL LABORATORIES CO., LTD. URL support@mbl.co.jp, TEL

2 References 1) Yin, F., et al., Stem Cells (214) In press. 2) Levine, J. E. et al., Blood 119, (212) 3) Harris, A. C. et al., Blood 119, (212) 4) Ferrara, J. L., et al., Blood 118, (211) ) Chen, C. Y., et al., Clin. Biochem. 38, 2-2 (2) 6) Motoo, Y., et al., Dig. Dis. Sci. 44, (1999) 7) Iovanna, J. L., et al., Gastroenterology 16, (1994) 8) Dusetti, N. J., et al., Genomics 19, (1994)

3 Calibration curve 2. PAP1 Calibration curve PAP1 Calibration Curve A STD conc. ng/ml Conc. (ng/ml) References data 1. Intra-assay ng/ml CV% 2. Detection limit (n=8) LOB (Limit of Blank) LOQ (Limit of Quantification) ng/ml 3.41 ng/ml 3. Reactivity with established cancer cell line culture supernatant * These samples were diluted (1:2) with culture medium prior to the assay. PAP1 conc. Of culture sup. from established cancer cell line ng/ml ) PAP1 concentration ( T24 MCF7 T47D OUMS-23 LoVo WiDr GCIY GT3TKB HGC27 KATO3 MKN1 MKN4 MKN74 OCUM-1 SCH A427 A49 ChaGo-K-1 DMS 114 H1793 H217 H226 H38 H2 H22 H96 LC174 LC319 LU61 PC-14 PC14PE6 PC-9 SK-LU-1 SK-MES-1 KLM1 MIA PaCa-2 PK-1 PK-9 22Rv1 PC-3 HUVEC 293T RPMI DMEM Bladder Breast Colon Gastric Lung Pancreatic Prostate n.c.

4 4. Normal range (9% reference interval) 3 PAP1 Serum form healthy samples (n=8) ng/ml * Serum samples were diluted (1:1) prior to the assay. 2 2 n ng/ml

5 CODE No. 323 For Research Use Only, Not for use in diagnostic procedures. Ab-Match Assembly Human PAP1 (REG3α) Kit 特徴 Ab-Match Assembly Human PAP1 (REG3α) Kit は Ab-Match Universal Kit(MBL; code no. 31) と組み合わせてご使用いただくことでサンドイッチ ELISA 法にて試料中のヒト PAP1 を特異的に測定することができますこのセットで 96 穴マイクロプレート 1 枚分の測定が可能です本製品は研究用試薬ですヒトの体内に用いたり診断の目的に使用しないでくださいキット構成 1 96 wells Name Materials Quantity Dilution ratio PAP1 standard Recombinant human PAP1* 2 µl 1 vial 1:1 1:32 or 1:64 Anti-PAP1 capture antibody Anti-PAP1 detection antibody Mouse anti-pap1 monoclonal antibody (IgG) (% Glycerol/PBS) Biotin conjugated rabbit anti-pap1 polyclonal antibody 6 µl 1 vial 1:2 11 µl 1 vial 1:11 Streptavidin conj. peroxidase Streptavidin conjugated peroxidase (SA-HRP) 11 µl 1 vial 1:11 * 濃度はラベルに記載してあります凍結融解を避けるため溶解後は分注してください注 ) キットの構成品は室温 (2-3 C) に戻してから使用してください操作法 Coating step 希釈した capture antibody を 1 に分注 2-8 C で一晩静置技術資料や関連情報はホームページからご利用頂けます Blocking step 2 回洗浄 Blocking Agent を 2 に分注室温で 1 時間静置 Sample Well の内容物を完全に除去調整した standard 及びサンプルを 1 µl ずつ各 well に分注室温で 1 時間静置 Detection antibody 4 回洗浄希釈した detection antibody を 1 に分注室温で 1 時間静置 SA-HRP step 4 回洗浄希釈した SA-HRP を 1 に分注室温で 3 分間静置保存と有効 -2 ~ -3 C にて保存してください有効期間は出荷後 12 ヶ月です関連製品 31 Ab-Match Universal Kit 参考文献 1) Yin, F., et al., Stem Cells (214) In press. 2) Levine, J. E. et al., Blood 119, (212) 3) Harris, A. C. et al., Blood 119, (212) 4) Ferrara, J. L., et al., Blood 118, (211) ) Chen, C. Y., et al., Clin. Biochem. 38, 2-2 (2) 6) Motoo, Y., et al., Dig. Dis. Sci. 44, (1999) 7) Iovanna, J. L., et al., Gastroenterology 16, (1994) 8) Dusetti, N. J., et al., Genomics 19, (1994) 4 回洗浄 Solution を 1 に分注室温で 3 分間静置 Reading Stop Solution を 1 µl ずつ各 well に分注 4 nm にて吸光度を測定 MEDICAL & BIOLOGICAL LABORATORIES CO., LTD. URL support@mbl.co.jp, TEL

6 標準曲線例 2. PAP1 Calibration curve PAP1 Calibration Curve A STD conc. ng/ml Conc. (ng/ml) 参考データ 1. 同時再現性 2. 検出感度 (n=8) ng/ml CV% LOB (Limit of Blank) LOQ (Limit of Quantification).24 ng/ml 3.41 ng/ml 3. 株化癌細胞培養上清との反応性 * 培養上清は培養液を用いて 2 倍希釈しました PAP1 conc. Of culture sup. from established cancer cell line ng/ml ) PAP1 concentration ( T24 MCF7 T47D OUMS-23 LoVo WiDr GCIY GT3TKB HGC27 KATO3 MKN1 MKN4 MKN74 OCUM-1 SCH A427 A49 ChaGo-K-1 DMS 114 H1793 H217 H226 H38 H2 H22 H96 LC174 LC319 LU61 PC-14 PC14PE6 PC-9 SK-LU-1 SK-MES-1 KLM1 MIA PaCa-2 PK-1 PK-9 22Rv1 PC-3 HUVEC 293T RPMI DMEM Bladder Breast Colon Gastric Lung Pancreatic Prostate n.c.

7 4. 標準範囲 (9% reference interval) n1 1 PAP1 Serum form healthy samples (n=8) ng/ml * 血清は 1 倍希釈しました ng/ml

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$(Microsoft Word _Ab-Match_Uni\201i2012_Sep\201j.doc)$ For Research Use Only, Not for use in diagnostic procedures. Ab-Match Universal kit For technical material or related information, please refer to https://ruo.mbl.co.jp/product/cancer/abmatch.html. Advantage