Human NKX2.5 Antibody

R&D Systems | Catalog # AF2444

R&D Systems

Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human

Cited:

Human, Mouse

Applications

Validated:

Immunohistochemistry, Western Blot

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Flow Cytometry, Immunocytochemistry, RNA immunoprecipitation

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all NKX2.5 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human NKX2.5
Gln24-Arg137
Accession # P52952

Specificity

Detects human NKX2.5 in direct ELISAs and Western blots.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human NKX2.5 Antibody

Detection of Human NKX2.5 by Western Blot

Detection of Human NKX2.5 by Western Blot

EOMES programs hESCs into functional CMs at high efficiency. a Illustration of growth factor-mediated and optimized EOMES induction-based cardiac differentiation protocols. Bottom right: Immunoblot validating doxycycline-dependent EOMES expression in a transgenic EOMESKO/E.TET-ON hESC line. b Typical yields of hESC-CMs (left, flow cytometry) and NKX2.5 expression (right, immunoblot) obtained with the two protocols (day 10). c Immunostainings 21 days after the initiation of EOMES induction. Weak perinuclear ANP staining is typical in overall MLC2v-positive hESC-CMs. Scale bars: 25 (top) and 50 µm (bottom). d Acceleration and slowdown of spontaneous beat rates in pCMs following exposure to 10 µM isoprenaline and 10 µM propranolol, respectively, on multielectrode arrays. e Microarray-based time course analysis comparing the indicated protocols and cell lines. RESCUE cells carry an inducible EOMES transgene on EOMESKO HuES6 background. Underlying data are from Supplementary Data 2 Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29382828), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human NKX2.5 by Immunocytochemistry/Immunofluorescence

Detection of Human NKX2.5 by Immunocytochemistry/Immunofluorescence

Contextual requirements of EOMES-mediated CM programming. a DOX dose dependency of the EOMES TET-ON protocol using the WTE.TET-ON hESC line. Top panel: Immunostains at 1.5 wk. Scale bar: 100 µm. Bottom: Flow cytometry analysis. Numbers indicate average percentages of cTnT-positive CMs from 3–6 experiments per condition. b CM programming by EOMES necessitates suppression of autocrine WNT signaling from the third day of transgene induction (qPCR data, n = 2). c DOX dose-dependent CM programming using an independent WTE.TET-ON hiPSC line. The data shows immunostaining (scale bar: 100 µm), RT-qPCR (n = 4), and FACS analysis (n = 3) performed at 1 wk of differentiation. The asymmetrical shape of the DOX titration data with this line is in part due to the incomplete repression of SOX2 at 0.05 µg/ml, which caused overgrowth of the cultures with neural precursors (also see Supplementary Fig. 3f). Error bars: s.e.m. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29382828), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human NKX2.5 by Immunocytochemistry/Immunofluorescence

Detection of Human NKX2.5 by Immunocytochemistry/Immunofluorescence

EOMES knockout hESCs fail to differentiate into cardiomyocytes. a Immunoblot confirming EOMES expression and its absence in WT and KO cells, respectively, at the cardiac mesoderm stage of directed differentiation. b EOMES KO cells fail to express the early cardiomyocyte marker NKX2.5 following exposure to a directed differentiation protocol. Scale bar: 50 µm. c EOMES KO cells show a general failure in markedly upregulating essential pan-cardiac genes (qPCR data, n = 3; error bars: s.e.m.). For each gene, the expression level in the highest expressing sample (“max” = WT in all three cases) is set to 1. d Differentiation of EOMES KO hESCs under signaling factor-assisted, non-cardiac mesoderm/endoderm-permissive culture conditions. Top: EOMES KO cells fail to express alpha -fetoprotein following Activin A-assisted endodermal induction. Scale bar: 50 µm. Bottom: Non-cardiac mesodermal differentiation competence of EOMES KO hESCs. KO cells were differentiated using varied meso and endoderm-permissive induction conditions. The data denote transformed p-values of annotation terms enriched in gene sets upregulated in EOMES KO samples compared to undifferentiated hESCs or differentiated WT controls (cutoffs: 10 or 3-fold, respectively). Underlying data: Supplementary Data 1 Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29382828), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human NKX2.5 Antibody

Application
Recommended Usage

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human heart

Western Blot

0.1 µg/mL
Sample: Recombinant Human NKX2.5

Reviewed Applications

Read 2 reviews rated 4 using AF2444 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: NKX2.5

The homeobox transcriptional factor NKX2.5 (Homeobox protein NK-2 homolog E) specifies cardiac and visceral mesoderm and plays an essential role in heart development.

Long Name

NK2 Homeobox 5

Alternate Names

CSX1, NKX2-5, NKX2E

Entrez Gene IDs

1482 (Human)

Gene Symbol

NKX2-5

UniProt

P52952

Additional NKX2.5 Products

Product Documents for Human NKX2.5 Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human NKX2.5 Antibody

For research use only

Citations for Human NKX2.5 Antibody

Customer Reviews for Human NKX2.5 Antibody (2)

4 out of 5
2 Customer Ratings
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  • Name: Anonymous
    Application: Immunofluorescence
    Sample Tested: See PMID 24123507
    Species: Mouse
    Verified Customer | Posted 01/07/2015
  • Name: Anonymous
    Application: Immunofluorescence
    Sample Tested: See PMID 24123507
    Species: Other
    Verified Customer | Posted 01/06/2015

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