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51133
TACSTD2/TROP2 (E8Y8S) Rabbit mAb (Alexa Fluor® 647 Conjugate)
Antibody Conjugates
Monoclonal Antibody

TACSTD2/TROP2 (E8Y8S) Rabbit mAb (Alexa Fluor® 647 Conjugate) #51133

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Flow Cytometry Image 1: TACSTD2/TROP2 (E8Y8S) Rabbit mAb (Alexa Fluor® 647 Conjugate)
Flow cytometric analysis of U-118 MG cells (blue, negative) and MDA-MB-468 cells (green, positive) using TACSTD2/TROP2 (E8Y8S) Rabbit mAb (Alexa Fluor® 647 Conjugate) (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control (Alexa Fluor® 647 Conjugate) #2985 (dashed lines).
To Purchase # 51133S
Product # Size Price
51133S
100 µl  (50 tests) $ 323

Supporting Data

REACTIVITY H
SENSITIVITY Endogenous
MW (kDa)
Source/Isotype Rabbit IgG

Application Key:

  • WB-Western Blot
  • IP-Immunoprecipitation
  • IHC-Immunohistochemistry
  • ChIP-Chromatin Immunoprecipitation
  • IF-Immunofluorescence
  • F-Flow Cytometry
  • E-P-ELISA-Peptide

Species Cross-Reactivity Key:

  • H-Human
  • M-Mouse
  • R-Rat
  • Hm-Hamster
  • Mk-Monkey
  • Vir-Virus
  • Mi-Mink
  • C-Chicken
  • Dm-D. melanogaster
  • X-Xenopus
  • Z-Zebrafish
  • B-Bovine
  • Dg-Dog
  • Pg-Pig
  • Sc-S. cerevisiae
  • Ce-C. elegans
  • Hr-Horse
  • All-All Species Expected

Product Description

This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 647 fluorescent dye and tested in-house for direct flow cytometric analysis in human cells. This antibody is expected to exhibit the same species cross-reactivity as the unconjugated TACSTD2/TROP2 (E8Y8S) Rabbit mAb #47866.

Product Usage Information

Application Dilution
Flow Cytometry 1:50

Storage

Supplied in PBS (pH 7.2), less than 0.1% sodium azide and 2 mg/ml BSA. Store at 4°C. Do not aliquot the antibody. Protect from light. Do not freeze.

Protocol

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Flow Cytometry, Live Cell Protocol for Directly Conjugated Antibodies

A. Solutions and Reagents

NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.

  1. 1X Phosphate Buffered Saline (PBS): To prepare 1 L 1X PBS: add 100 ml 10X PBS (#12528) to 900 ml water, mix.
  2. Antibody Dilution Buffer: Purchase ready-to-use Flow Cytometry Antibody Dilution Buffer (#13616), or prepare a 0.5% BSA PBS buffer by dissolving 0.5 g Bovine Serum Albumin (BSA) (#9998) in 100 ml 1X PBS. Store at 4°C.

NOTE: When including fluorescent cellular dyes in your experiment (including viability dyes, DNA dyes, etc.), please refer to the dye product page for the recommended protocol. Visit www.cellsignal.com for a full listing of cellular dyes validated for use in flow cytometry.

B. Immunostaining

NOTE: Count cells using a hemocytometer or alternative method.

NOTE: If using whole blood, lyse red blood cells and wash by centrifugation prior to Immunostaining.

NOTE: Optimal centrifugation conditions will vary depending upon cell type and reagent volume. Generally, 150-300g for 1-5 minutes will be sufficient to pellet the cells.

  1. Aliquot desired number of cells into tubes or wells. (Generally, 5x105 to 1x106 cells per assay.)
  2. Pellet cells by centrifugation and remove supernatant.
  3. Resuspend cells in 100 µl of diluted primary antibody, prepared in Antibody Dilution Buffer at a recommended dilution or as determined via titration.
  4. Incubate for 30 min to 1 hr on ice. Protect from light.
  5. Wash by centrifugation in Antibody Dilution Buffer. Discard supernatant. Repeat.
  6. Resuspend cells in 200-500 µl of Antibody Dilution Buffer and analyze on flow cytometer.

posted June 2017

revised June 2020

Protocol Id: 1504

Specificity / Sensitivity

TACSTD2/TROP2 (E8Y8S) Rabbit mAb (Alexa Fluor® 647 Conjugate) recognizes endogenous levels of total TACSTD2/TROP2 protein.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Val90 of human TACSTD2/TROP2 protein.

Background

TROP2 is a transmembrane glycoprotein encoded by gene TACSTD2 (tumor associated calcium signal transducer 2). TROP2 was first discovered as a biomarker of invasive trophoblast cells and later reported in many types of cancer cells as well as in various organs during development, and adult stem cells during homeostasis (1,2). TROP2 has an extracellular domain with EGF thyroglobulin type-1 repeats, a transmembrane domain, and a short cytoplasmic tail with a HIKE domain containing a PIP2 binding site and PKC phosphorylation site (Ser303) (1-4). TROP2 functions by regulating multiple signaling pathways including interaction of extracellular domain with integrin beta1 to regulate FAK signaling, association of its transmembrane domain with Claudin-1 and Claudin-7 for tight junction formation, as well as regulation of intracellular calcium release by its PIP2 binding and activation of the ERK/MAPK pathway (1,2,5-8). All of these functions are important for its role in tumor proliferation, metastasis, and invasion (1,2). PKC can phosphorylate TROP2 at Ser303; the phosphorylation changes the cytoplasmic tail conformation and further promotes its signaling (9). TROP2 can be activated through intramembrane proteolysis first by TACE, followed by further cleavage by Presenilin 1 and Presenilin 2. The proteolysis process is required for its role in tumor cell proliferation (10,11).
  1. McDougall, A.R. et al. (2015) Dev Dyn 244, 99-109.
  2. Shvartsur, A. and Bonavida, B. (2015) Genes Cancer 6, 84-105.
  3. El Sewedy, T. et al. (1998) Int J Cancer 75, 324-30.
  4. Linnenbach, A.J. et al. (1993) Mol Cell Biol 13, 1507-15.
  5. Trerotola, M. et al. (2013) Cancer Res 73, 3155-67.
  6. Trerotola, M. et al. (2015) Oncotarget 6, 14318-28.
  7. Nakatsukasa, M. et al. (2010) Am J Pathol 177, 1344-55.
  8. Cubas, R. et al. (2010) Mol Cancer 9, 253.
  9. Pavšič, M. et al. (2015) Sci Rep 5, 10324.
  10. Stoyanova, T. et al. (2012) Genes Dev 26, 2271-85.
  11. Ju, X. et al. (2016) Cancer Res 76, 6723-34.

Pathways & Proteins

Explore pathways + proteins related to this product.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST's products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST's Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.
This product is provided under an intellectual property license from Life Technologies Corporation. The transfer of this product is conditioned on the buyer using the purchased product solely in research conducted by the buyer, excluding contract research or any fee for service research, and the buyer must not (1) use this product or its components for (a) diagnostic, therapeutic or prophylactic purposes; (b) testing, analysis or screening services, or information in return for compensation on a per-test basis; or (c) manufacturing or quality assurance or quality control, and/or (2) sell or transfer this product or its components for resale, whether or not resold for use in research. For information on purchasing a license to this product for purposes other than as described above, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or [email protected]