Vitamin D Binding protein Recombinant Rabbit Monoclonal Antibody [JM10-36]
cat.: ET1703-09
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: WB, IF-Cell, IF-Tissue, IHC-P, FC
Clonality: Monoclonal
Clone number: JM10-36
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 53 kDa
Isotype: IgG
Immunogen: Synthetic pertide within Human Vitamin D Binding protein aa 270-319 / 474.
Positive control: Human plasma lysates, human pancreas tissue, Hela, HepG2, SKOV-3.
Subcellular location: Secreted.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  FC
1:2,000
1:50-1:200
1:50-1:200
1:50-1:200
1:50-1:100
Uniprot #: SwissProt: P02774 Human
Alternative names: DBP DBP/GC GC Gc globulin Gc-globulin GRD3 Group specific component Group specific component vitamin D binding protein Group-specific component hDBP VDB VDBG VDBP Vitamin D binding alpha globulin Vitamin D-binding protein VTDB_HUMAN
Images
ET1703-09_1.jpg Fig1: Western blot analysis of Vitamin D Binding protein on human plasma lysates with Rabbit anti-Vitamin D Binding protein antibody (ET1703-09) at 1/2,000 dilution.

Lysates/proteins at 20 µg/Lane.

Predicted band size: 53 kDa
Observed band size: 53 kDa

Exposure time: 20 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1703-09) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1703-09_2.jpg Fig2: Immunohistochemical analysis of paraffin-embedded human pancreas tissue with Rabbit anti-Vitamin D Binding protein antibody (ET1703-09) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1703-09) at 1/1,000 dilution for 1 hour 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.
ET1703-09_3.jpg Fig3: ICC staining of Vitamin D Binding protein in Hela cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1703-09, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1703-09_4.jpg Fig4: ICC staining of Vitamin D Binding protein in HepG2 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1703-09, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1703-09_5.jpg Fig5: ICC staining of Vitamin D Binding protein in SKOV-3 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1703-09, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1703-09_6.jpg Fig6: Flow cytometric analysis of Vitamin D Binding protein was done on HepG2 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1703-09, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.