GLi3 (GLI-Kruppel Family Member GLI3)

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.\n\nApplications: \nSuitable for use in ELISA, Immunocytochemistry, and Western Blot. Other applications have not been tested.\n\nRecommended Dilution:\nWestern 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. \nImmunocytochemistry: 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. \nDirect 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.\n\nOptimal dilutions should be determined by each laboratory for each application.\n\nStorage and Stability:\nLyophilized powder may be stored at 4 degrees C for short-term only. Reconstitute to nominal volume by adding sterile PBS with 40-50% glycerol and store at -20 degrees C. Reconstituted product is stable for 12 months at -20 degrees 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.

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SPECIFICATIONS

Catalog Number

G2035-54

Size

100ug

Applications

ELISA, ICC, WB

Hosts

Goat

Reactivities

Hum, Mouse

Form

Supplied as a lyophilized powder from a 0.2um filtered solution in PBS with 5% trehalose. Reconstitute with sterile PBS, 40-50% glycerol. If 0.5ml is used, the concentration will be 0.2mg/ml.

P Type

Pab

Purity

Purified by human GLI-3 affinity chromatography.

Isotype

IgG

Additional Info

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.