Detection of Human PRDM16 by Western Blot.
Western blot shows lysates of K562 human chronic myelogenous leukemia cell line. PVDF Membrane was probed with 1 µg/mL of Sheep Anti-Human/Mouse PRDM16 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6295) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for PRDM16 at approximately 170 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.
PRDM16 in Mouse Embryo.
PRDM16 was detected in immersion fixed frozen sections of mouse embryo (E13.5) using Sheep Anti-Human/Mouse PRDM16 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6295) at 10 µg/mL overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (orange, upper panel; Catalog # NL010) and counterstained with DAPI (blue, lower panel). Specific staining was localized to the trigeminal ganglion. View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.
Preparation and Storage
Sterile PBS to a final concentration of 0.2 mg/mL.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & 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.
PRDM16 (PR [PRDI-BF1 and RIZ] domain containing protein 16; also MEL-1) is a 170 kDa member of the PR Domain family of proteins. It is a transcriptional regulator expressed in the embryo, and is reported to participate in the maintenance of both neuronal and hematopoietic progenitor stem cells populations, and to preferentially promote the development of brown fat from adipomyocyte precursors. The generation of brown fat is likely due to suppression of muscle-specific factors. Mouse PRDM16 is 1275 amino acids (aa) in length. It contains one SET domain (aa 85-208) followed by ten C2H2 type Zn finger motifs (aa 230-1030). There are multiple potential isoform variants that likely vary from 150-170 kDa in size. One isoform shows a deletion of aa 1232-1250, a second isoform shows a three aa substitution for aa 1174-1275, and a third isoform possesses an alternative start site at Met21, coupled to a deletion of aa 1196-1133. Over aa 537-688, mouse PRDM16 shares 81% and 95% aa identity with human and rat PRDM16, respectively.
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We have 1 review tested in 1 species: Mouse.
We have 1 review tested in 1 application: Western Blot.
Western Blot: Human/Mouse PRDM16/MEL1 Antibody [AF6295].
Adipose tissue,Brown adipose tissue
Other Experimental Details
Other Experimental Details
Samples were nuclear extracts from brown adipose and subcutaneous adipose tissue (20ug protein loaded). Primary antibody (1:1,000) was detected with HRP-anti sheep (1:10,000) then amplified with Licor streptavidin 800 (1:10,000). Near infrared fluorescence gave much better signal-to-noise than chemiluminescence.