PRODUCT CODE: ET1610-63

Recombinant DKK1 Monoclonal Antibody (ET1610-63)

  • Recombinant

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Rat

Western blot analysis of DKK1 on MCF-7 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1610-63, 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.
  • Western blot analysis of DKK1 on MCF-7 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1610-63, 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.
  • ICC staining of DKK1 in Hela 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 (ET1610-63, 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).
  • ICC staining of DKK1 in HepG2 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 (ET1610-63, 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).
  • ICC staining of DKK1 in NCCIT 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 (ET1610-63, 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 human uterus tissue using anti-DKK1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) 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 (ET1610-63, 1/50) 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 DKK1 was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1610-63, 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 DKK1 on MCF-7 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1610-63, 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.

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Rat

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

Recombinant DKK1 Monoclonal Antibody (ET1610-63)

Immunogen

Recombinant protein

Host

Rabbit

Positive Control

MCF-7 cell lysates, Hela, HepG2, NCCIT, human uterus tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

SC06-86

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

38 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:1,000

  • ICC/IF

  • 1:50-1:500

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

TARGET

UNIPROT #

PROTEIN NAME

DKK1

SYNONYMS

Dickkopf 1 antibody; Dickkopf 1 homolog antibody; Dickkopf 1 like antibody; Dickkopf homolog 1 antibody; Dickkopf like protein 1 antibody; Dickkopf related protein 1 antibody; Dickkopf WNT signaling pathway inhibitor 1 antibody; Dickkopf-1 antibody; Dickkopf-related protein 1 antibody; DKK 1 antibody; Dkk-1 antibody; Dkk1 antibody; DKK1_HUMAN antibody; hDkk 1 antibody; hDkk-1 antibody; SK antibody

SEQUENCE SIMILARITIES

Belongs to the dickkopf family.

TISSUE SPECIFICITY

Placenta.

SUBCELLULAR LOCATION

Secreted.

FUNCTION

The Wnt genes are a group of well conserved, cysteine-rich secreted glycoproteins that are required for numerous developmental processes including embryogenesis, asymmetric cell division and central nervous system (CNS) patterning. Wnt association with the seven membrane spanning receptor frizzled activates dishevelled, which downregulates glycogen synthase kinase (GSK) through serine phosphorylation, causing the accumulation of b-catenin and subsequent regulation of developmentally significant Wnt |target genes. The Dickkopf family of secreted inhibitors of Wnt signaling ensures proper morphological development by antagonizing different stages of the Wnt cascade. Dkk-1 (Dickkopf-1) acts upstream of b-catenin and dishevelled to inhibit Wnt signaling. Dkk-1 is a 266-amino acid (human), secreted protein that contains a 31-residue N-terminal signal peptide, 2 cysteine rich domains, and a putative carboxy terminal N-glycosylation site. Human Dkk-1 transcripts are abundantly present in fetal kidney, adult placenta and adult prostate. Putative cis regulatory elements upstream of the Dkk-1 start site include p53, Sp1, MyoD, STAT, Oct-1/2, C/EBP-a, C/EBP-b, GATA-1, GATA-2 and GATA-3.