NKX3.1 Antibody

Western blot shows lysates of LNCaP human prostate cancer cell line. PVDF Membrane was probed with 0.1 µg/mL of Goat Anti-Human NKX3.1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6080) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for NKX3.1 at approximately 38 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.

NKX3.1 was detected in immersion fixed LNCaP human prostate cancer cell line using Goat Anti-Human NKX3.1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6080) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Activation of beta -catenin pathway in LNCaP cells overexpressing δ-catenin(A) Comparison of levels of δ-catenin in overexpressing LNCaP clones and other PCa cell lines. Full length protein is depicted by arrow. (B) Levels of catenin proteins in δ-catenin overexpressing LNCaP clone (designated as OE1b) are shown. Gradient loading of total protein for OE1b clone was used to illustrate an increase in expression of each protein (right panel). Quantification graphs showing protein levels normalized to GAPDH level (arbitrary units, a.u.) are at the bottom. (C) Characterization of nuclear (NE) and cytoplasmic (CE) levels of beta -catenin. Nuclear and cytoplasmic protein was isolated, beta -catenin was detected by Western blotting using specific antibody. TBP and GAPDH were used as loading control of nuclear and cytoplasmic protein respectively. Normalized level of beta -catenin in each compartment is shown at the bottom. (D and E) Characterization of levels of proteins, downstream targets of Wnt/ beta -catenin pathway (D), and androgen regulated genes AR and NKX3.1 (E) by Western blotting. Corresponding quantification is shown. Three clones (OE3a, OR1b, OE1c) overexpressing δ-catenin at various levels were examined. Total amount of 30ug of protein was used in each experiment, unless otherwise specified (in B). SDS-PAGE and Western blot conditions as in Figure 2. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29849951), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: WB, ICC/IF
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

NKX3.1 Antibody Summary

• Immunogen: E. coli-derived recombinant human NKX3.1
  Met1-Pro123
  Accession # Q99801
• Specificity: Detects human NKX3.1 in direct ELISAs and Western blots. In direct ELISAs, less than 1% cross-reactivity with recombinant human (rh) NKX2.2, rhNKX6.1, and rhBAP31 is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: NKX3-1
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 5-15 ug/mL
  • Western Blot 0.1 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for NKX3.1 Antibody

• BAPX2
• BAPX2homeobox protein Nkx-3.1
• Homeobox protein NK-3 homolog A
• NK homeobox (Drosophila), family 3, A
• NK3 homeobox 1
• NK3 transcription factor homolog A
• NK3 transcription factor related, locus 1 (Drosophila)
• NK3 transcription factor related, locus 1
• NKX3.1
• NKX3.1NKX3
• NKX3-1
• NKX3A
• NKX3ANK homeobox, family 3, A

Background

NKX3.1 (Homeobox protein NK-3 homolog A) is a 32-38 kDa member of the NK-3 homeobox family of transcription factors. It is expressed in testis and prostatic epithelium, and appears to be regulated by both testosterone and estrogen (but not progesterone). NKX3.1 apparently promotes prostate gland development, and acts as a tumor suppressor/transcriptional repressor by interacting with HDAC-1 and increasing p53 acetylation and half-life. Human NKX3.1 is 234 amino acids (aa) in length. It contains a DNA binding homeodomain (aa 124-183) and phosphorylation sites at Ser185 and Ser196 that regulate protein turnover. There are four splice variants that show changes N-terminal to the homeobox domain. Deletions of aa 8-56, 13-87, 15-83 and 40-83 are all reported. Over aa 1-123, human NKX3.1 shares 50% aa identity with mouse NKX3.1.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for NKX3.1 Antibody (AF6080)(1)

Publication using AF6080

• Zhang P, Schaefer-Klein J, Cheville JC et al. Frequently rearranged and overexpressed ?-catenin is responsible for low sensitivity of prostate cancer cells to androgen receptor and ?-catenin antagonists. Oncotarget 2018-05-11 [PMID: 29849951]

Bioinformatics

• Gene Symbol: NKX3-1
• Uniprot:
  • Q99801(Human)