{"product_id":"pab-exatecan-igg-labeling-kit-ua079063","title":"PAB-Exatecan IgG Labeling Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\n\u003cbr\u003e\u003ctable style=\"width: 100%; height: auto;\"\u003e\u003ctbody\u003e\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eStability \u0026amp; Storage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp\u003e\u003cspan\u003eDry ice transport. Store protected from light at −20°C or lower; for long-term storage (\u0026gt;3 months), we recommend −80°C. Refer to the reagent vial label for the expiration date. After first use, it is recommended to store the reagent at 4°C for short-term use (within 2 weeks). For longer storage, aliquot into single-use portions and store protected from light at −20°C or lower, avoiding repeated freeze-thaw cycles.\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/div\u003e\u003ch4\u003eBackground\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp\u003e\u003cspan\u003ePAB protein (Protein of Antibody Binding, MW 25 kDa) contains the IgG Fc-binding domain derived from Protein G and includes a cysteine residue at its C-terminus. Under reducing conditions, the C-terminal cysteine of PAB is converted into a reduced thiol group. Through appropriate coupling reactions, toxins or dyes, including pH-sensitive dyes, can be specifically attached to this reduced thiol group. Such site-specific conjugation does not interfere with the specific binding between PAB and the IgG Fc region. PAB conjugated with toxins or dyes, including pH-sensitive dyes, can serve as a universal binding reagent for human or mouse IgG antibodies, enabling the study of antibody internalization or phagocytosis in target cells.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003eExatecan (also known as irinotecan or irinotecan hydrochloride) is a water-soluble derivative of camptothecin (CPT). It exhibits potent antitumor activity and, in tumor cells, induces apoptosis by inhibiting DNA topoisomerase I. It is one of the commonly used cytotoxic payloads in antibody–drug conjugates (ADCs).\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003eThe PAB-Exatecan IgG labeling kit includes the PAB-Exatecan reagent, which is a complex where MC-Val-Cit-PAB-Exatecan is site-specifically conjugated to PAB via a reductive thiol group at the C-terminus of PAB. The conjugation between PAB and Exatecan is achieved using the widely used cleavable linker MC-VC-PABC, which contains a maleimide spacer, the Val-Cit dipeptide sequence as a substrate for cathepsin, and the self-cleaving PABC spacer. PAB-Exatecan can serve as a universal binding reagent for human or mouse IgG antibodies, and by detecting the cytotoxic effect of the antibody-PAB-Exatecan complex on target cells, it reflects the endocytic or phagocytic activity of the antibody. The PAB-Exatecan IgG labeling reagent employs a unique method to remove unlabeled free MC-Val-Cit-PAB-Exatecan, making the process simple and rapid—antibody labeling can be completed within 30 minutes. The labeled antibody does not require purification and can be directly used in subsequent endocytosis or phagocytosis assays. Compared with commonly used conjugated toxins such as DT3C, the PAB-Exatecan reagent has a smaller molecular weight, thereby reducing the potential impact of PAB-IgG conjugation on antibody function. Additionally, the VC-PABC linker better mimics the actual application of ADCs.\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eComponents\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan\u003ePAB-\u003c\/span\u003e\u003cspan\u003eExatecan\u003c\/span\u003e\u003cspan\u003eIgG Labeling Kit\u003c\/span\u003e\u003cspan\u003eSpecifications are as follows. The amount of labeled IgG is calculated based on a molar ratio of 3:1, using PAB with a molecular weight of 25 kDa and IgG antibody with a molecular weight of 150 kDa.\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003e\u003c\/span\u003e\u003cspan\u003e.\u003c\/span\u003e\u003c\/p\u003e\n\u003ctable style=\"border-collapse:collapse;width:auto;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eSpecifications\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eReagent\u003c\/span\u003e\u003cspan style=\"font-weight:bold\"\u003eVolume\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eReagent\u003c\/span\u003e\u003cspan style=\"font-weight:bold\"\u003eConcentration (PAB)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eLabeled IgG\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003e50 μg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"color:#000000\"\u003e2x50\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eμL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"color:#000000\"\u003e250\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eμ\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eg\/mL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"color:#000000\"\u003e50\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eμg\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/table\u003e\n\u003cbr\u003e\n\u003c\/div\u003e\u003ch4\u003eProtocol\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan\u003eAccording to\u003c\/span\u003e\u003cspan\u003ePAB MW 25KDa\u003c\/span\u003e\u003cspan\u003eand\u003c\/span\u003e\u003cspan\u003eIgG\u003c\/span\u003e\u003cspan\u003eantibody\u003c\/span\u003e\u003cspan\u003eMW 150KDa\u003c\/span\u003e\u003cspan\u003ecalculation,\u003c\/span\u003e\u003cspan\u003eIgG\u003c\/span\u003e\u003cspan\u003eand\u003c\/span\u003e\u003cspan\u003ePAB-Exatecan\u003c\/span\u003e\u003cspan\u003elabeled in a molar ratio of\u003c\/span\u003e\u003cspan\u003e3\u003c\/span\u003e\u003cspan\u003e:\u003c\/span\u003e\u003cspan\u003e1\u003c\/span\u003e\u003cspan\u003ecan meet the needs of most experiments, and the molar ratio can also be optimized according to specific experimental requirements, up to a maximum of\u003c\/span\u003e\u003cspan\u003e6\u003c\/span\u003e\u003cspan\u003e:\u003c\/span\u003e\u003cspan\u003e1\u003c\/span\u003e\u003cspan\u003e, which can enhance the detection signal intensity. The following experiment uses\u003c\/span\u003e\u003cspan\u003eTrastuzumab\u003c\/span\u003e\u003cspan\u003e \u003c\/span\u003e\u003cspan\u003eand\u003c\/span\u003e\u003cspan\u003ePAB-Exatecan\u003c\/span\u003e\u003cspan\u003ecombined in a molar ratio of\u003c\/span\u003e\u003cspan\u003e3\u003c\/span\u003e\u003cspan\u003e:\u003c\/span\u003e\u003cspan\u003e1\u003c\/span\u003e\u003cspan\u003eto form a complex, and assesses its cytotoxic effect on\u003c\/span\u003e\u003cspan\u003eHER-2\u003c\/span\u003e\u003cspan\u003eexpressing\u003c\/span\u003e\u003cspan\u003eSK-BR-3\u003c\/span\u003e\u003cspan\u003ecells to reflect the antibody's internalization efficiency.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e1)\u003c\/span\u003e\u003cspan\u003eIn a\u003c\/span\u003e\u003cspan\u003e96\u003c\/span\u003e\u003cspan\u003ewell white transparent cell culture plate, seed\u003c\/span\u003e\u003cspan\u003eMcCoy's 5A\u003c\/span\u003e\u003cspan\u003e \u003c\/span\u003e\u003cspan\u003e(10% FBS)\u003c\/span\u003e\u003cspan\u003eculture medium at\u003c\/span\u003e\u003cspan\u003e100\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003el\u003c\/span\u003e\u003cspan\u003eper well,\u003c\/span\u003e\u003cspan\u003e4,000\u003c\/span\u003e\u003cspan\u003eSK-BR-3\u003c\/span\u003e\u003cspan\u003ecells\u003c\/span\u003e\u003cspan\u003e,\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eincubate overnight in a\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003e37\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003e℃\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003e \u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003e5% CO\u003c\/span\u003e\u003csub\u003e\u003cspan style=\"color:#000000\"\u003e2\u003c\/span\u003e\u003c\/sub\u003e\u003cspan style=\"color:#000000\"\u003e \u003c\/span\u003e\u003cspan\u003eincubator\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e2)\u003c\/span\u003e\u003cspan\u003ePrepare\u003c\/span\u003e\u003cspan\u003e10x\u003c\/span\u003e\u003cspan\u003etest concentrations\u003c\/span\u003e\u003cspan\u003eTrastuzumab-PAB-Exatecan\u003c\/span\u003e\u003cspan\u003eComplex: As required by the experiment\u003c\/span\u003e\u003cspan\u003eTake an appropriate amount\u003c\/span\u003e\u003cspan\u003eOpti-MEM I\u003c\/span\u003e\u003cspan\u003eDilute in culture medium\u003c\/span\u003e\u003cspan\u003eTrastuzumab\u003c\/span\u003e\u003cspan\u003eto\u003c\/span\u003e\u003cspan\u003e50\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003eg\/mL\u003c\/span\u003e\u003cspan\u003e, then according to\u003c\/span\u003e\u003cspan\u003e10\u003c\/span\u003e\u003cspan\u003e:\u003c\/span\u003e\u003cspan\u003e1\u003c\/span\u003e\u003cspan\u003eadd\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003e250\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eμ\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eg\/mL\u003c\/span\u003e\u003cspan\u003e \u003c\/span\u003e\u003cspan\u003eof\u003c\/span\u003e\u003cspan\u003ePAB-Exatecan\u003c\/span\u003e\u003cspan\u003eto a final concentration of\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003e22.7\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eμ\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eg\/mL\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003e, for example,\u003c\/span\u003e\u003cspan\u003e30\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003eL\u003c\/span\u003e\u003cspan\u003eof\u003c\/span\u003e\u003cspan\u003e50\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003eg\/mL Trastuzumab\u003c\/span\u003e\u003cspan\u003eadd\u003c\/span\u003e\u003cspan\u003e3\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eL\u003c\/span\u003e\u003cspan\u003e250\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eμ\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eg\/mL\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eof\u003c\/span\u003e\u003cspan\u003e \u003c\/span\u003e\u003cspan\u003ethe\u003c\/span\u003e\u003cspan\u003ePAB-Exatecan\u003c\/span\u003e\u003cspan\u003e, thus\u003c\/span\u003e\u003cspan\u003ethe concentrations of\u003c\/span\u003e\u003cspan\u003eTrastuzumab\u003c\/span\u003e\u003cspan\u003eand\u003c\/span\u003e\u003cspan\u003ePAB-Exatecan\u003c\/span\u003e\u003cspan\u003e(10x)\u003c\/span\u003e\u003cspan\u003eare approximately\u003c\/span\u003e\u003cspan\u003e300 nM\u003c\/span\u003e\u003cspan\u003eand\u003c\/span\u003e\u003cspan\u003e900 nM\u003c\/span\u003e\u003cspan\u003e, mix well by pipetting, and incubate at room temperature\u003c\/span\u003e\u003cspan\u003efor 30 minutes\u003c\/span\u003e\u003cspan\u003e.\u003c\/span\u003e\u003cspan\u003eIt is recommended that after the first use of\u003c\/span\u003e\u003cspan\u003ePAB-Exatecan, it be stored short-term\u003c\/span\u003e\u003cspan\u003e(2 weeks) at\u003c\/span\u003e\u003cspan\u003e4\u003c\/span\u003e\u003cspan\u003e℃, and for long-term storage, aliquot according to single-use amounts and store protected from light\u003c\/span\u003e\u003cspan\u003eat -20\u003c\/span\u003e\u003cspan\u003e℃\u003c\/span\u003e\u003cspan\u003eor lower\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e3)\u003c\/span\u003e\u003cspan\u003eusing\u003c\/span\u003e\u003cspan\u003eOpti-MEM I\u003c\/span\u003e\u003cspan\u003eThe prepared medium was used to perform a 3X serial dilution of the above-prepared\u003c\/span\u003e\u003cspan\u003eTrastuzumab\u003c\/span\u003e\u003cspan\u003e \u003c\/span\u003e\u003cspan\u003eand\u003c\/span\u003e\u003cspan\u003ePAB-Exatecan\u003c\/span\u003e\u003cspan\u003ecomplex, resulting in a total of\u003c\/span\u003e\u003cspan\u003e8\u003c\/span\u003e\u003cspan\u003econcentration levels.\u003c\/span\u003e\u003cspan\u003e4)\u003c\/span\u003e\u003cspan\u003eRemove the\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003eSK-BR-3\u003c\/span\u003e\u003cspan\u003eexperimental cell plate, and add\u003c\/span\u003e\u003cspan\u003e10\u003c\/span\u003e\u003cspan\u003eμL of the gradient concentrations of the above-prepared\u003c\/span\u003e\u003cspan\u003eTrastuzumab-PAB-Exatecan\u003c\/span\u003e\u003cspan\u003ecomplex to each well. Based on\u003c\/span\u003e\u003cspan\u003eTrastuzumab\u003c\/span\u003e\u003cspan\u003e, the highest final concentration is calculated to be\u003c\/span\u003e\u003cspan\u003e30 nM.\u003c\/span\u003e\u003cspan\u003e,\u003c\/span\u003e\u003cspan\u003e3x\u003c\/span\u003e\u003cspan\u003edilutions, totaling\u003c\/span\u003e\u003cspan\u003e8\u003c\/span\u003e\u003cspan\u003econcentration levels.\u003c\/span\u003e\u003cspan\u003e5)\u003c\/span\u003e\u003cspan\u003eIncubate at\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e37\u003c\/span\u003e\u003cspan\u003e℃\u003c\/span\u003e\u003cspan\u003eand\u003c\/span\u003e\u003cspan\u003e \u003c\/span\u003e\u003cspan\u003e5%\u003c\/span\u003e\u003cspan\u003eCO2\u003c\/span\u003e\u003cspan\u003ein an incubator for\u003c\/span\u003e\u003cspan\u003e4\u003c\/span\u003e\u003cspan\u003edays; it is recommended to replenish with\u003c\/span\u003e\u003cspan\u003e20\u003c\/span\u003e\u003cspan\u003eμ\u003c\/span\u003e\u003cspan\u003eL McCoy's 5A\u003c\/span\u003e\u003cspan\u003e(\u003c\/span\u003e\u003cspan\u003e10% FBS\u003c\/span\u003e\u003cspan\u003e) culture medium after\u003c\/span\u003e\u003cspan\u003e3\u003c\/span\u003e\u003cspan\u003edays.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e6)\u003c\/span\u003e\u003cspan\u003ePerform\u003c\/span\u003e\u003cspan\u003ecell viability\u003c\/span\u003e\u003cspan\u003eassays;\u003c\/span\u003e\u003cspan\u003e \u003c\/span\u003e\u003cspan\u003efor example, you can use the\u003c\/span\u003e\u003cspan\u003eUA-Glo® Luminescent Cell Viability Assay\u003c\/span\u003e\u003cspan\u003e(\u003c\/span\u003e\u003cspan\u003eUA070103\u003c\/span\u003e\u003cspan\u003e) to measure cell viability.\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eGuidelines\u003c\/h4\u003e\u003cdiv\u003eDo not mix different batches.\u003cbr\u003eDo not alter the volume of the test reagent without rigorous validation.\u003cbr\u003eFor research use only.\u003c\/div\u003e","brand":"UA BIOSCIENCE","offers":[{"title":"50μg","offer_id":67433046999115,"sku":"UA079063-50μg","price":865.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/AntBioImage_be5310fb-b375-4c6d-a397-ad708bb5f960.png?v=1794250815","url":"https:\/\/www.antbioinc.com\/products\/pab-exatecan-igg-labeling-kit-ua079063","provider":"AntBio","version":"1.0","type":"link"}