PRODUCT CODE: ET1610-2

Nanog Recombinant Rabbit Monoclonal Antibody [SC05-70] (ET1610-2)

  • Recombinant

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Mouse

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Western blot analysis of Nanog on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1610-2, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: F9 cell lysate<br />
Lane 2: mouse testis tissue lysate
  • Western blot analysis of Nanog on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1610-2, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: F9 cell lysate<br />
Lane 2: mouse testis tissue lysate
  • ICC staining of Nanog in F9 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1610-2, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded mouse testis tissue using anti-Nanog antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-2, 1/200)  for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Flow cytometric analysis of Nanog was done on NCCIT cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1610-2, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Western blot analysis of Nanog on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1610-2, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: F9 cell lysate
Lane 2: mouse testis tissue lysate

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Mouse

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

Nanog Recombinant Rabbit Monoclonal Antibody [SC05-70] (ET1610-2)

Immunogen

Recombinant protein

Host

Rabbit

Positive Control

F9 cell lysate, mouse testis tissue lysate, F9, NCCIT.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

SC05-70

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A affinity purified.

MOLECULAR WEIGHT

37 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:1,000-1:2,000

  • ICC/IF

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

TARGET

UNIPROT #

PROTEIN NAME

Nanog

SYNONYMS

Embryonic stem cell specific homeobox protein (Nanog) antibody; ENK antibody; FLJ12581 antibody; hNanog antibody; Homeobox protein NANOG antibody; Homeobox transcription factor Nanog antibody; homeobox transcription factor Nanog-delta 48 antibody; NANOG antibody; Nanog homeobox antibody; NANOG_HUMAN antibody

SEQUENCE SIMILARITIES

Belongs to the Nanog homeobox family.

TISSUE SPECIFICITY

Expressed in testicular carcinoma and derived germ cell tumors (at protein level). Expressed in fetal gonads, ovary and testis. Also expressed in ovary teratocarcinoma cell line and testicular embryonic carcinoma. Not expressed in many somatic organs and oocytes.

DEVELOPMENTAL STAGE

Expressed in embryonic stem (ES) and carcinoma (EC) cells. Expressed in inner cell mass (ICM) of the blastocyst and gonocytes between 14 and 19 weeks of gestation (at protein level). Not expressed in oocytes, unfertilized oocytes, 2-16 cell embryos and early morula (at protein level). Expressed in embryonic stem cells (ES). Expression decreases with ES differentiation.

SUBCELLULAR LOCATION

Nucleus.

FUNCTION

Nanog (from “Tir Na Nog,” the mythologic Celtic land of the ever young) is a divergent homeodomain protein that directs pluripotency and differentiation of undifferentiated embryonic stem cells. Nanog mRNA is present in pluripotent mouse and human cell lines and absent from differentiated cells. Human Nanog protein shares 52% overall amino acid identity with the mouse protein and 85% identity in the homeodomain. Human Nanog maps to gene locus 12p13.31, whereas mouse Nanog maps to gene loci 6 F2. Murine embryonic Nanog expression is detected in the inner cell mass of the blastocyst. High levels of human Nanog expression have been detected by Northern analysis in the undifferentiated NTERA-2 cl.D1 embryonal carcinoma cell line.

CITATIONS

  • Zhu, Xinfeng et al.

    All-Trans Retinoic Acid-Induced Deficiency of the Wnt/β-Catenin Pathway Enhances Hepatic Carcinoma Stem Cell Differentiation. | PloS One [2015]