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Anti-Human TRA-1-60 Antibody, Clone TRA-1-60R

Mouse monoclonal IgM antibody against human, rhesus, rabbit TRA-1-60 (podocalyxin)

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Anti-Human TRA-1-60 Antibody, Clone TRA-1-60R

Mouse monoclonal IgM antibody against human, rhesus, rabbit TRA-1-60 (podocalyxin)

From: 250 USD
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Mouse monoclonal IgM antibody against human, rhesus, rabbit TRA-1-60 (podocalyxin)
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Overview

The TRA‐1‐60R antibody reacts with TRA-1-60 (Podocalyxin), a > 200 kDa pluripotent stem cell-specific protein expressed on the surface of undifferentiated human embryonic stem (ES) and induced pluripotent stem (iPS) cells, embryonal carcinoma (EC) cells and embryonic germ (EG) cells, as well as rhesus monkey ES cell lines. A soluble form of TRA-1-60 has been detected in serum of patients with embryonal carcinoma. The epitope, which is lost upon cell differentiation, contains sialic acid and is associated with a large-molecular-mass transmembrane protein named podocalyxin. Though sialylated, the epitope recognized by the TRA‐1‐60R antibody is resistant to treatment with neuraminidase.

This antibody clone has been verified for labeling human ES and iPS cells grown in TeSR™-E8™ (Catalog #05940), mTeSR™1 (Catalog #85850), and TeSR™2 (Catalog #05860), and for purity assessments of cells isolated with EasySep™ kits, including EasySep™ Human ES/iPS Cell TRA-1-60 Positive Selection Kit (Catalog #18166; partial blocking may be observed) and EasySep™ hESC/hiPSC SSEA-4 Positive Selection Kit (Catalog #18165).
Subtype
Primary Antibodies
Target Antigen
TRA-1-60 (Podocalyxin)
Alternative Names
Podocalyxin, TRA-1
Reactive Species
Human, Rabbit, Rhesus
Conjugation
Alexa Fluor 488, Biotin, PE, Unconjugated
Host Species
Mouse
Cell Type
Pluripotent Stem Cells
Species
Human, Non-Human Primate, Other
Application
Cell Isolation, Flow Cytometry, Immunocytochemistry, Immunofluorescence, Immunoprecipitation, Western Blotting
Area of Interest
Stem Cell Biology
Clone
TRA-1-60R
Gene ID
5420
Isotype
IgM, kappa

Data Figures

Data for Alexa Fluor® 488-Conjugated

Figure 1. Data for Alexa Fluor® 488-Conjugated

(A) Flow cytometry analysis of human ES cells (filled histogram) or HT1080 fibrosarcoma cells (negative control, dashed line histogram) labeled with Anti-Human TRA-1-60 Antibody, Clone TRA-1-60R, Alexa Fluor® 488. Labeling of human ES cells with Mouse IgM, kappa Isotype Control Antibody, Clone MM-30, Alexa Fluor® 488 is shown (Catalog #60069AD, solid line histogram).
(B) Human ES cells were cultured in mTeSR™1 on BD Matrigel™-coated glass slides, then fixed and stained with Anti-Human TRA-1-60 Antibody, Clone TRA-1-60R, Alexa Fluor® 488. Inset shows cells labeled with Mouse IgM, kappa Isotype Control Antibody, Clone MM-30, Alexa Fluor® 488.
(C) Flow cytometry analysis of human iPS cells labeled with Anti-Human TRA-1-60 Antibody, Clone TRA-1-60R, Alexa Fluor® 488 (filled histogram) or Mouse IgM, kappa Isotype Control Antibody, Clone MM-30, Alexa Fluor® 488 (open histogram).
(D) Human iPS cells were cultured in mTeSR™1 on BD Matrigel™-coated glass slides, then fixed and stained with Anti-Human TRA-1-60 Antibody, Clone TRA-1-60R, Alexa Fluor® 488. Inset shows cells labeled with Mouse IgM, kappa Isotype Control Antibody, Clone MM-30, Alexa Fluor® 488.

Data for Biotin-Conjugated

Figure 2. Data for Biotin-Conjugated

Flow cytometry analysis of human ES cells (filled histogram) or HT1080 fibrosarcoma cells (negative control, dashed line histogram) labeled with Anti-Human TRA-1-60 Antibody, Clone TRA-1-60R, Biotin followed by streptavidin (SAV) APC (filled histogram). Labeling human ES cells with a mouse IgM, kappa biotin isotype control antibody followed by SAV APC is shown (solid line histogram).

Data for PE-Conjugated

Figure 3. Data for PE-Conjugated

(A) Flow cytometry analysis of human embryonic stem (ES) cells (filled histogram) or HT1080 fibrosarcoma cells (negative control, dashed line histogram) labeled with Anti-Human TRA-1-60 Antibody, Clone TRA-1-60R, PE. Labeling of human ES cells with Mouse IgM, kappa Isotype Control Antibody, Clone MM-30, PE is shown (Catalog #60069PE, solid line histogram).
(B) Human ES cells were cultured in mTeSR™1 on BD Matrigel™-coated glass slides, then fixed and stained with Anti-Human TRA-1-60 Antibody, Clone TRA-1-60R, PE. Inset shows cells labeled with Mouse IgM, kappa Isotype Control Antibody, Clone MM-30, PE.
(C) Flow cytometry analysis of human iPS cells labeled with Anti-Human TRA-1-60 Antibody, Clone TRA-1-60R, PE (filled histogram) or Mouse IgM, kappa Isotype Control Antibody, Clone MM-30, PE (open histogram).
(D) Flow cytometry analysis of human ES cells isolated with the EasySep™ Human ES/iPS Cell TRA-1-60 Positive Selection Kit from a mixed population of ES cells and HT1080 fibrosarcoma cells and labeled with Anti-Human TRA-1-60 Antibody, Clone TRA-1-60R, PE. Histograms show labeling of the starting population containing ~5% ES cells (Start) and the isolated cells (Isolated). Labeling with Mouse IgM, kappa Isotype Control Antibody, Clone MM-30, PE is shown in the bottom panel (open histogram).

Data for Unconjugated

Figure 4. Data for Unconjugated

(A) Flow cytometry analysis of human ES cells (filled histogram) or HT1080 fibrosarcoma cells (negative control, dashed line histogram) labeled with Anti-Human TRA-1-60 Antibody, Clone TRA-1-60R, followed by goat anti-mouse IgG, FITC. Labeling of human ES cells with Mouse IgM, kappa Isotype Control Antibody, Clone MM-30 (Catalog #60069) followed by goat anti-mouse IgG, FITC is shown (solid line histogram).
(B) Human ES cells were cultured in mTeSR™1 on BD Matrigel™-coated glass slides, then fixed and stained with Anti-Human TRA-1-60 Antibody, Clone TRA-1-60R, followed by goat anti-mouse IgG, FITC. Inset shows cells labeled with Mouse IgM, kappa Isotype Control Antibody, Clone MM-30, followed by goat anti-mouse IgG, FITC.
(C) Flow cytometry analysis of human iPS cells labeled with Anti-Human TRA-1-60 Antibody, Clone TRA-1-60R, followed by goat anti-mouse IgG, FITC (filled histogram) or Mouse IgM, kappa Isotype Control Antibody, Clone MM-30, followed by goat anti-mouse IgG, FITC (open histogram).
(D) Western blot analysis of denatured/reduced cell lysates from human ES cells (lane 1) or HT1080 fibrosarcoma cells (negative control, lane 2) with Anti-Human TRA-1-60 Antibody, Clone TRA-1-60R.

Protocols and Documentation

Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.

Document Type
Product Name
Catalog #
Lot #
Language
Document Type
Product Information Sheet
Product Name
Anti-Human TRA-1-60 Antibody, Clone TRA-1-60R, Alexa Fluor® 488
Catalog #
60064AD, 60064AD.1
Lot #
All
Language
English
Document Type
Product Information Sheet
Product Name
Anti-Human TRA-1-60 Antibody, Clone TRA-1-60R, PE
Catalog #
60064PE, 60064PE.1
Lot #
All
Language
English
Document Type
Product Information Sheet
Product Name
Anti-Human TRA-1-60 Antibody, Clone TRA-1-60R
Catalog #
60064
Lot #
All
Language
English
Document Type
Safety Data Sheet
Product Name
Anti-Human TRA-1-60 Antibody, Clone TRA-1-60R, Alexa Fluor® 488
Catalog #
60064AD, 60064AD.1
Lot #
All
Language
English
Document Type
Safety Data Sheet
Product Name
Anti-Human TRA-1-60 Antibody, Clone TRA-1-60R, PE
Catalog #
60064PE, 60064PE.1
Lot #
All
Language
English
Document Type
Safety Data Sheet
Product Name
Anti-Human TRA-1-60 Antibody, Clone TRA-1-60R
Catalog #
60064
Lot #
All
Language
English

Applications

This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.

Research Area
Workflow Stages

Resources and Publications

Educational Materials (3)

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Publications (1)

CaMKII inhibition rectifies arrhythmic phenotype in a patient-specific model of catecholaminergic polymorphic ventricular tachycardia. Di Pasquale E et al. Cell death & disease 2013

Abstract

Induced pluripotent stem cells (iPSC) offer a unique opportunity for developmental studies, disease modeling and regenerative medicine approaches in humans. The aim of our study was to create an in vitro 'patient-specific cell-based system' that could facilitate the screening of new therapeutic molecules for the treatment of catecholaminergic polymorphic ventricular tachycardia (CPVT), an inherited form of fatal arrhythmia. Here, we report the development of a cardiac model of CPVT through the generation of iPSC from a CPVT patient carrying a heterozygous mutation in the cardiac ryanodine receptor gene (RyR2) and their subsequent differentiation into cardiomyocytes (CMs). Whole-cell patch-clamp and intracellular electrical recordings of spontaneously beating cells revealed the presence of delayed afterdepolarizations (DADs) in CPVT-CMs, both in resting conditions and after $\$-adrenergic stimulation, resembling the cardiac phenotype of the patients. Furthermore, treatment with KN-93 (2-[N-(2-hydroxyethyl)]-N-(4methoxybenzenesulfonyl)]amino-N-(4-chlorocinnamyl)-N-methylbenzylamine), an antiarrhythmic drug that inhibits Ca(2+)/calmodulin-dependent serine-threonine protein kinase II (CaMKII), drastically reduced the presence of DADs in CVPT-CMs, rescuing the arrhythmic phenotype induced by catecholaminergic stress. In addition, intracellular calcium transient measurements on 3D beating clusters by fast resolution optical mapping showed that CPVT clusters developed multiple calcium transients, whereas in the wild-type clusters, only single initiations were detected. Such instability is aggravated in the presence of isoproterenol and is attenuated by KN-93. As seen in our RyR2 knock-in CPVT mice, the antiarrhythmic effect of KN-93 is confirmed in these human iPSC-derived cardiac cells, supporting the role of this in vitro system for drug screening and optimization of clinical treatment strategies.
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Legal Statement:

Alexa Fluor is a registered trademark of Life Technologies Corporation. Antibodies conjugated to Alexa Fluor® are licensed for internal research use only and sale is expressly conditioned on the buyer not using the antibody for manufacturing, performing a service or medical test, or otherwise generating revenue. For use other than research, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@lifetech.com.

Quality Statement:

PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED. FOR ADDITIONAL INFORMATION ON QUALITY AT STEMCELL, REFER TO WWW.STEMCELL.COM/COMPLIANCE.

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