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Figure 1 Immunoflouorescence Validation of NANOG in HeLa Cells</strong><br>Immunofluorescent analysis of PFA-fixed HeLa cells labeling NANOG with 5585 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/1000 dilution (red) and DAPI staining (blue). Alpha tubulin was stained with anti-alpha tubulin antibody following by goat anti-mouse IgG secondary antibody (green). Images were captured with confocal microscopy.
<strong>Figure 2 WB Validation in HeLa Cells</strong><br>Loading: 15 μg of lysate. Antibodies: NANOG 5585, 2 μg/mL , 1 h incubation at RT in 8% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10,000 dilution.
<strong>Figure 3 Western Blot Validation in Mouse and Rat Testis </strong><br> Loading: 15 μg of lysates per lane. Antibodies: NANOG 5585, 2 μg/mL, 1h incubation at RT in 8% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10,000 dilution.
<strong>Figure 4 Immunohistochemistry Validation of NANOG in Human Testis </strong><br> Immunohistochemical analysis of paraffin-embedded human testis tissue using anti-NANOG antibody (5585) at 5 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
<strong>Figure 5 Immunohistochemistry Validation of NANOG in Mouse Testis </strong><br> Immunohistochemical analysis of paraffin-embedded mouse testis tissue using anti-NANOG antibody (5585) at 2 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
<strong>Figure 6 Immunohistochemistry Validation of NANOG in Rat Testis </strong><br> Immunohistochemical analysis of paraffin-embedded rat testis tissue using anti-NANOG antibody (5585) at 2 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
Figure 1 Immunoflouorescence Validation of NANOG in HeLa Cells
Immunofluorescent analysis of PFA-fixed HeLa cells labeling NANOG with 5585 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/1000 dilution (red) and DAPI staining (blue). Alpha tubulin was stained with anti-alpha tubulin antibody following by goat anti-mouse IgG secondary antibody (green). Images were captured with confocal microscopy.
Trial Size

NANOG Antibody

CATALOG NUMBER: 5585

Clonality:
Polyclonal
Tested Applications:
ELISA, IHC-P, WB, ICC/IF
Host Species:
Rabbit
Species Reactivity:
Human, Mouse, Rat
Conjugate:
Unconjugated
Specifications
Host Species:
Rabbit
Species Reactivity:
Human, Mouse, Rat
Homology:
Predicted species reactivity based on immunogen sequence: Bovine: (89%), Chicken: (94%)
Immunogen:
NANOG antibody was raised against a 19 amino acid synthetic peptide near the center of human NANOG.
The immunogen is located within amino acids 130 - 180 of NANOG.
Conjugate:
Unconjugated
Tested Applications:
ELISA, IHC-P, WB, ICC/IF
Application Note:
WB: 1-2 μg/mL; ICC/IF: 20 μg/mL; IHC: 1-5 μg/mL.
Antibody validated: Western Blot in human, mouse and rat samples; Immunohistochemistry in human, mouse and rat samples and Immunofluorescence in human samples. All other applications and species not yet tested.
Positive Control 1:
Cat. No. 1416 – Mouse Testis Tissue Lysate
Positive Control 2:
Cat. No. 1476 – Rat Testis Tissue Lysate
Predicted Molecular Weight:
Predicted: 35 kD Observed: 35 kD
Isoforms:
Human NANOG has 2 isoforms, isoform 1 (305aa, 34.6kD) and isoform 2 (289aa, 32.8kDa). Mouse NANOG has 2 isoforms, isoform 1 (305aa, 34.2kD) and isoform 2 (280aa, 31.9kD). Rat NANOG has 1 isoform (311aa, 35.1kD). 5585 can detect NANOG in human, mouse, and rat.
Purification:
NANOG Antibody is affinity chromatography purified via peptide column.
Clonality:
Polyclonal
Isotype:
IgG
physical-state:
Liquid
Buffer:
NANOG Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Storage Conditions:
NANOG antibody can be stored at 4°C for three months and -20°C, stable for up to one year. As with all antibodies care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures.
Ncbi Official Symbol:
NANOG
Additional Names:
NANOG Antibody: Homeobox protein NANOG, Homeobox transcription factor Nanog, hNanog
Protein Accession Number:
EAW88651
Protein Gi Number:
119609057
Ncbi Gene Id Number:
79923
User Note:
Optimal dilutions for each application to be determined by the researcher.
Background:
NANOG Antibody: Expression of NANOG is required for the maintenance of pluripotency in epiblast and embryonic stem (ES) cells as well as for the ability to maintain ES self-renewal independently of LIF/Stat3. The role of NANOG in embryonic development suggested that it might be useful in the creation of stem cells that might be useful in cell replacement therapies in the treatment of several degenerative diseases. Artificial stem cells, termed induced pluripotent stem (iPS) cells, can be created by expressing POU5F1 (also known as Oct-4), another germline-specific transcription factor, and the transcription factors Sox2, Klf4 and Lin28 along with c-Myc in mouse fibroblasts. More recently, experiments have demonstrated that iPS cells could be generated using expression plasmids expressing NANOG, Sox2, KlfF4 and c-Myc, eliminating the need for virus introduction, thereby addressing a safety concern for potential use of iPS cells in regenerative medicine.
Background Reference 1:
Chambers I, Colby D, Robertson M, et al. Functional expression cloning of Nanog, a pluripotency sustaining factor in embryonic stem cells. Cell2003; 113:643-55.
Background Reference 2:
Mitsui K, Tokuzowa Y, Itoh H, et al. The homeoprotein Nanog is required for maintenance of pluripotency in mouse epiblast and ES cells. Cell2003; 113:631-42.
Background Reference 3:
Carpenter MK, Rosler E, and Rao MS. Characterization and differentiation of human embryonic stem cells. Cloning Stem Cells2003; 5:79-88.
Background Reference 4:
Scholer HR, Ruppert S, Suzuki N, et al. New type of POU domain in germ line-specific protein Oct-2. Nature1990; 344:435-9.

FOR RESEARCH USE ONLY

For additional information, visit ProSci’s Terms & Conditions Page.

Disclaimer:
Optimal dilutions/concentrations should be determined by the end user. The information provided is a guideline for product use. This product is for research use only.

CATALOG NUMBER:

5585

List Size:
0.02 mg, 0.1 mg

List Price:

Price range: $99.00 through $445.00

Datasheet
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