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GLI3 Antibody
SKU: orb345478
Description
Images & Validation
−Item 1 of 5
| Tested Applications | ELISA, IF, IHC, Multiplex Assay, WB |
|---|---|
| Dilution range | ELISA: 1:6,000 - 1:30,000, IHC: 0.5 mg/ml - 5 µg/ml, WB: 1:500 - 1:2,000 |
| Reactivity | Human |
| Application Notes |
Key Properties
−| Antibody Type | Primary Antibody |
|---|---|
| Host | Rabbit |
| Clonality | Polyclonal |
| Isotype | IgG |
| Immunogen | This affinity purified antibody was produced from monospecific rabbit serum by repeated immunizations with a synthetic peptide corresponding to an internal region near amino acids 30-60 of human Gli-3 protein. |
| Purity | This affinity-purified antibody is directed against human Gli-3 protein. The product was affinity purified from monospecific antiserum by immunoaffinity purification. A BLAST analysis was used to suggest cross reactivity with Gli-3 from human, chimpanzee, squirrel monkey, Xenopus laevis, chicken, dog and quail based on 100% sequence homology with the immunogen. Reactivity is also expected against homologues from mouse (94%) and rat (88%) based on partial homology. Reactivity with Gli-3 from other sources is not known. |
| Conjugation | Unconjugated |
Storage & Handling
−| Storage | Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use. |
|---|---|
| Form/Appearance | Liquid (sterile filtered) |
| Buffer/Preservatives | 0.01% (w/v) Sodium Azide |
| Concentration | 1.0 mg/mL |
| Dry Ice Shipping | Please note: This product requires shipment on dry ice. A dry ice surcharge will apply. |
| Disclaimer | For research use only |
Alternative Names
−Rabbit anti-GLI-3 antibody, Transcriptional activator GLI3, Gli 3, GLI3 form of 190 kDa, GLI3 form of 83 kDa
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Immunofluorescence Microscopy of Rabbit anti-Gli-3 antibody. Tissue: MCF-7 cell. Antigen retrieval: not required. Primary antibody: Gli-3 antibody and Anti-alpha-Tubulin at 5 µg/ml for 1 h at RT. Secondary antibody: Fluorescein secondary antibody at 1:10000 for 45 min at RT. Localization: Gli-3 is nuclear. Staining: Gli-3 staining as red fluorescent signal and Anti-alpha-Tubulin staining as green fluorescent signal using STED.
Immunofluorescence Microscopy of Rabbit anti-Gli-3 antibody. Tissue: MCF-7 cell. Antigen retrieval: not required. Primary antibody: Gli-3 antibody and Anti-alpha-Tubulin at 5 µg/ml for 1 h at RT. Secondary antibody: Fluorescein secondary antibody at 1:10000 for 45 min at RT. Localization: Gli-3 is nuclear. Staining: Image (1) shows alpha-Tubulin staining as green fluorescent signal. Image (2) shows Gli-3 staining as red fluorescent signal and Images (3) shows both antibodies fluorescing using STED microscopy.
Immunohistochemistry of Rabbit anti-Gli-3 antibody. This image tissue: human glioblastoma. Specific staining was also noted in tissue from adrenal, brain, glioblastoma, colon, heart, kidney, lung, liver, skeletal muscle, ovary, pancreas, placenta, skin, spleen, stomach, testes, thymus, thyroid, tonsil and uterus. Fixation: formalin fixed paraffin embedded. Antigen retrieval: not required. Primary antibody: Gli-3 antibody at 0.625 µg/ml for 1 h at RT. Secondary antibody: Peroxidase rabbit secondary antibody at 1:10000 for 45 min at RT. Localization: Gli-3 is nuclear and smooth muscle. Staining: Gli-3 as precipitated red signal with hematoxylin purple nuclear counterstain.
Western Blot of Rabbit anti-Gli-3 antibody. Lane 1: 50 kDa molecular weight marker. Lane 2: 293T cells transfected with CrkL-Flag. Lane 3: 293T cells transfected with human Gli-3. Load: 35 µg per lane. Primary antibody: Gli-3 antibody at 1:400 for overnight at 4°C. Secondary antibody: IRDye800™ rabbit secondary antibody at 1:10000 for 45 min at RT. Block: 5% BLOTTO overnight at 4°C. Predicted/Observed size: 170-190 kDa for hGli-3. Other band(s): Non specific background ~60kDa.
Western Blot of Rabbit anti-Gli-3 antibody. Lane 1: human brain whole cell lysate. Lane 2: human lung whole cell lysate. Lane 3: human spleen whole cell lysate. Lane 4: mouse brain whole cell lysate (p/n orb348715). Lane 5: mouse lung whole cell lysate (p/n orb692725). Load: 20 µg per lane. Primary antibody: Gli-3 antibody at 1:500 for overnight at 4°C. Secondary antibody: IRDye800™ rabbit secondary antibody at 1:10000 for 45 min at RT. Block: 5% BLOTTO overnight at 4°C. Predicted/Observed size: Isoforms at ~170-190kDa and ~80kDa. Lane 2 shows what may be truncated Gli-3 (~80kDa). Other band(s): The strong band at ~50 kDa is unknown.
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WB
Western Blot (IB, immunoblot)
IHC
Immunohistochemistry
IF
Immunofluorescence
ELISA
Enzyme-linked Immunosorbent Assay (EIA)
GLI3 Antibody (orb345478)
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