Zinc Finger Protein GLI1 (GLI1) (C-Term), (AA 800-850) antibody

Details for Product No. ABIN153487
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Antigen
Epitope
C-Term, AA 800-850
(3), (3), (2), (2), (2), (1), (1), (1), (1), (1), (1), (1)
Reactivity
Human
(45), (19), (15), (15), (13), (6), (1), (1)
Host
Rabbit
(44), (7), (4), (2), (1)
Clonality
Polyclonal
Conjugate
Un-conjugated
(2), (2), (2), (1), (1), (1), (1), (1), (1), (1), (1)
Application
Western Blotting (WB), Chromatin Immunoprecipitation (ChIP), ELISA, Immunohistochemistry (IHC), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p))
(40), (19), (10), (7), (6), (6), (3), (2), (1)
Pubmed 9 references available
Quantity 0.1 mL
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Catalog No. ABIN153487
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Immunogen A synthetic peptide made toward the C-terminus portion of the human Gli1 protein (between residues 800-850)
Specificity This is directed against Gli1 from mouse brain. No reaction occurs with human or mouse Gli2 or Gli3.
Purification Affinity purified
Alternative Name GLI1
Background This antiserum was produced against a peptide corresponding to the carboxy-terminalregion of the mouse Gli-1 protein. This region is not conserved among other gli familymembers, namely Gli-2 and Gli-3. Gli was termed by Kinzler et al. (1987) as',glioma-associated oncogene', amplified in malignant gliomas. Analysis ofthe cloned gene demonstrates that the gene contains 5 repeats of zinc-fingersequences, which places gli in the family of Kruppel (Kr) zinc finger proteins. Northernanalysis reveals that GLI is expressed in embryonal carcinoma cells but not in mostadult tissue. GLI has been localized to 12q13-q14.3 by Southern blot analysis. Onmouse the gene is located on chromosome 10. In mice, 3 zinc finger transcriptionfactors, Gli-1, Gl-i2 and Gli-3, have been implicated in the transduction of Sonichedgehog (Shh) signal. In papillary epithelium, shh, gli1 and ptc all follow similarexpression patterns. Gli1 expression is central and probably sufficient for tumordevelopment in humans. Alternate Names: Anti-GLI antibody, Anti-Glioma associated oncogene antibody, Anti-Glioma associatedoncogene homolog 1 (zinc finger protein) antibody, Anti-Oncogene GLI antibody,Anti-Zinc finger protein GLI1 antibody.
Gene Symbol: GLI1
Gene ID 2735, 14632
UniProt P08151
Research Area Growth Factors
Application Notes This Gli1 antibody is useful for Western Blot and Immunohistochemistry-paraffin sections. Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (pH 6.0) is recommended.
Recommended dilutions: Chromatin Immunoprecipitation 1:10-1:500, ELISA 1:20000-1:100000, Immunohistochemistry 1:600, Immunohistochemistry-Paraffin 1:600, Western Blot 1:2000-1:10000
Protocol Western Blot protocol specific for Gli1 antibody Western Blot Protocol
1. Perform SDS-PAGE on samples to be analyzed, loading 40 µg of total protein per lane.
. Transfer proteins to membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
. Stain according to standard Ponceau S procedure (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
. Rinse the blot.
. Block the membrane using standard blocking buffer for at least 1 hour.
. Wash the membrane in wash buffer three times for 10 minutes each.
. Dilute primary antibody in blocking buffer and incubate 1 hour at room temperature.
. Wash the membrane in wash buffer three times for 10 minutes each.
. Apply the diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.
. Wash the blot in wash buffer three times for 10 minutes each (this step can be repeated as required to reduce background).
. Apply the detection reagent of choice in accordance with the manufacturers instructions.**Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2 %.Immunohistochemistry-Paraffin Embedded Sections Protocol Immunohistochemistry-Paraffin Embedded SectionsAntigen Unmasking:Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.
0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes. Staining:
. Wash sections in deionized water three times for 5 minutes each.
. Wash sections in wash buffer for 5 minutes.
. Block each section with 100-400 µL blocking solution for 1 hour at room temperature.
. Remove blocking solution and add 100-400 µL diluted primary antibody. Incubate overnight at 4C.
. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
. Add 100-400 µL biotinylated diluted secondary antibody. Incubate 30 minutes at room temperature.
. Remove secondary antibody solution and wash sections three times with wash buffer for 5 minutes each.
. Add 100-400 µL Streptavidin-HRP reagent to each section and incubate for 30 minutes at room temperature.
. Wash sections three times in wash buffer for 5 minutes each.
. Add 100-400 µL DAB substrate to each section and monitor staining closely.
. As soon as the sections develop, immerse slides in deionized water.
. Counterstain sections in hematoxylin.
. Wash sections in deionized water two times for 5 minutes each.
. Dehydrate sections.
. Mount coverslips.
Restrictions For Research Use only
Format Liquid
Concentration 1.05 mg/mL
Buffer PBS, 30 % glycerol, Sodium Azide
Preservative Sodium azide
Precaution of Use WARNING: Reagents contain sodium azide. Sodium azide is very toxic if ingested or inhaled. Avoid contact with skin, eyes, or clothing. Wear eye or face protection when handling. If skin or eye contact occurs, wash with copious amounts of water. If ingested or inhaled, contact a physician immediately. Sodium azide yields toxic hydrazoic acid under acidic conditions. Dilute azide-containing compounds in running water before discarding to avoid accumulation of potentially explosive deposits in lead or copper plumbing.
Handling Advice Avoid freeze-thaw cycles
Storage 4 °C
Storage Comment 4 °C short term. Aliquot and store at -20 °C long term.
Supplier Images
anti-Zinc Finger Protein GLI1 (GLI1) (C-Term), (AA 800-850) antibody anti-Zinc Finger Protein GLI1 (GLI1) (C-Term), (AA 800-850) antibody
General Kinzler, Ruppert, Bigner et al.: "The GLI gene is a member of the Kruppel family of zinc finger proteins." in: Nature, Vol. 332, Issue 6162, pp. 371-4, 1988 (PubMed).

Kinzler, Bigner, Bigner et al.: "Identification of an amplified, highly expressed gene in a human glioma." in: Science (New York, N.Y.), Vol. 236, Issue 4797, pp. 70-3, 1987 (PubMed).

Dahmane, Lee, Robins et al.: "Activation of the transcription factor Gli1 and the Sonic hedgehog signalling pathway in skin tumours." in: Nature, Vol. 389, Issue 6653, pp. 876-81, 1997 (PubMed).

Hall, Hooper, Finger: "Expression of sonic hedgehog, patched, and Gli1 in developing taste papillae of the mouse." in: The Journal of comparative neurology, Vol. 406, Issue 2, pp. 143-55, 1999 (PubMed).

Ghali, Wong, Green et al.: "Gli1 protein is expressed in basal cell carcinomas, outer root sheath keratinocytes and a subpopulation of mesenchymal cells in normal human skin." in: The Journal of investigative dermatology, Vol. 113, Issue 4, pp. 595-9, 1999 (PubMed).

Park, Bai, Platt et al.: "Mouse Gli1 mutants are viable but have defects in SHH signaling in combination with a Gli2 mutation." in: Development (Cambridge, England), Vol. 127, Issue 8, pp. 1593-605, 2000 (PubMed).

Wetmore, Eberhart, Curran: "The normal patched allele is expressed in medulloblastomas from mice with heterozygous germ-line mutation of patched." in: Cancer research, Vol. 60, Issue 8, pp. 2239-46, 2000 (PubMed).

Qualtrough, Buda, Gaffield et al.: "Hedgehog signalling in colorectal tumour cells: induction of apoptosis with cyclopamine treatment." in: International journal of cancer. Journal international du cancer, Vol. 110, Issue 6, pp. 831-7, 2004 (PubMed).

Breunig, Sarkisian, Arellano et al.: "Primary cilia regulate hippocampal neurogenesis by mediating sonic hedgehog signaling." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 105, Issue 35, pp. 13127-32, 2008 (PubMed).

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