Human GLI-3 is a 190kD transcription factor of the C2H2-type zinc-finger protein family.
GLI-3 appears to play a role in limb and brain development, probably through sonic hedgehog signaling.
GLI-3 is regulated by PKA-mediated phosphorylation.
Unphosphorylated full-length GLI-3 is a transcriptional activator, while phosphorylated GLI-3 is cleaved after amino acid 700 to generate an N-terminal 83kD transcriptional repressor.
Over the region used as immunogen, human GLI-3 is 95 % identical to the corresponding mouse and canine protein sequences.
Applications:Suitable for use in ELISA, Immunocytochemistry, and Western Blot.
Other applications have not been tested.
Recommended Dilution:Western Blot: 0.1-0.2ug/ml.
Used in conjunction with the appropriate secondary reagents, detects human and mouse GLI-3.
The detection limit for rhGLI-3 and rmGLI-3 is approximately 5 ng/lane under non- reducing and reducing conditions.
Immunocytochemistry: 10ug/ml detects human GLI-3 in HeLa cells.
Cells were fixed with PBS containing 4% paraformaldehyde and blocked with PBS containing 10% normal donkey serum, 0.1% Triton X-100, and 1% BSA.
After blocking, cells were incubated with diluted primary antibody followed by NorthernLights 557-coupled anti-goat IgG at room temperature in the dark.
Between each step, cells were washed with PBS containing BSA.
Direct ELISA: 0.5-1.0ug/ml.
Used in conjunctionthe appropriate secondary reagents, detects human and mouse GLI-3.
The detection limit for rhGLI-3 and rmGLI-3 is approximately 1 ng/well.
Optimal dilutions should be determined by each laboratory for each application.
Storage and Stability:Lyophilized powder may be stored at 4°C for short-term only.
Reconstitute to nominal volume by adding sterile PBS and store at -20°C.
Reconstituted product is stable for 12 months at -20°C.
For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Further dilutions can be made in assay buffer.
仕様
Size：100ug
Host：goat
Source Antibody：human
Grade：Affinity Purified
Purity：Purified by immunoaffinity chromatography.
Form：Supplied as a lyophilized powder in PBS, 5% trehalose. Reconstitute with 500ul sterile PBS.
Specificity：Recognizes human and mouse GLI-3 in the applications listed. In direct ELISAs and Western Blots, the antibody shows less than 1% crossreactivity with rhGLI-1 and rhGLI-2.
Isotype：IgG
Calc Applications Abbrev：E IC WB
Calc Crossreactivity：Hu Mo
Immunogen：E. coli-derived, recombinant human Glioma-associated oncogene homolog 3 (rhGLI-3; aa1-479)
