WB result of Osteocalcin Recombinant Rabbit mAb
Primary antibody: Osteocalcin Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: 293T transfected with empty vector whole cell lysate 20 µg
Lane 2: 293T transfected with Osteocalcin-Myc-His fusion protein whole cell lysate 20 µg
Secondary antibody: Goat Anti- rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 11 kDa
Observed MW: 13 kDa
Osteocalcin Recombinant Rabbit mAb (SDT-2311-37)
Osteocalcin Recombinant Rabbit mAb (SDT-2311-37)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | Osteocalcin |
| Synonyms | Bone Gla protein (BGP); Gamma-carboxyglutamic acid-containing protein; BGLAP |
| Immunogen | Synthetic Peptide |
| Location | Secreted |
| Accession | P02818 |
| Clone Number | SDT-2311-37 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, IHC-P, ICC |
| Reactivity | Hu, Ms, Rt |
| Purification | Protein A |
| Concentration | 0.25 mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300 |
| Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied |
Dilution
| application | dilution | species |
| WB | 1:1000-1:5000 | Hu |
| IHC-P | 1:2000 | Hu |
| ICC | 1:250 | Hu |
Background
Osteocalcin is a 49-residue, vitamin-K-dependent, γ-carboxyglutamic acid-rich protein synthesized exclusively by osteoblasts and odontoblasts that, once γ-carboxylated and incorporated into bone extracellular matrix where it regulates mineral maturation, can re-enter the circulation in an undercarboxylated form to act as a hormone that binds the GPCR6A receptor on pancreatic β-cells, adipocytes and myofibers to augment insulin secretion, enhance adiponectin expression and fuel muscle glucose uptake, thereby coupling skeletal remodeling to systemic energy metabolism and explaining why bone is an endocrine organ.
Picture
Picture
Western Blot
Immunohistochemistry
IHC shows positive staining in paraffin-embedded human Osteoblastoma. Anti-Osteocalcin antibody was used at 1/2000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human gastric cancer. Anti-Osteocalcin antibody was used at 1/2000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
Negative control: IHC shows negative staining in paraffin-embedded human kidney. Anti-Osteocalcin antibody was used at 1/2000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
Immunocytochemistry
ICC shows positive staining in Osteocalcin- myc-His transfected 293F cells (top panel) and negative staining in vector-transfected 293F cells (below panel). Anti-Osteocalcin antibody was used at 1/250 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 100% ice-cold methanol and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).
