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GLI2 antibody - middle region (ARP31885_T100)

100 ul
$249.00
In Stock

Conjugation Options

ARP31885_T100-FITC Conjugated

ARP31885_T100-HRP Conjugated

ARP31885_T100-Biotin Conjugated

Gene Symbol:
GLI2
NCBI Gene Id:
2736
Official Gene Full Name:
GLI family zinc finger 2
Protein Name:
Zinc finger protein GLI2
Swissprot Id:
P10070
Protein Accession #:
NP_084655
Nucleotide Accession #:
NM_030379
Alias Symbols:
HPE9, THP1, THP2
Replacement Item:
This antibody may replace item sc-20290 from Santa Cruz Biotechnology.
Description of Target:
GLI2 encodes a protein which belongs to the C2H2-type zinc finger protein subclass of the Gli family. Members of this subclass are characterized as transcription factors which bind DNA through zinc finger motifs. These motifs contain conserved H-C links. Gli family zinc finger proteins are mediators of Sonic hedgehog (Shh) signaling and they are implicated as potent oncogenes in the embryonal carcinoma cell. The protein encoded by this gene localizes to the cytoplasm and activates patched Drosophila homolog (PTCH) gene expression. It is also thought to play a role during embryogenesis. The encoded protein is associated with several phenotypes- Greig cephalopolysyndactyly syndrome, Pallister-Hall syndrome, preaxial polydactyly type IV, postaxial polydactyly types A1 and B.
Protein Size (# AA):
1258
Molecular Weight:
133kDa
Host:
Rabbit
Clonality:
Polyclonal
Purification:
Protein A purified
Application:
IHC, WB
Tissue Tool:
Find tissues and cell lines supported by DNA array analysis to express GLI2.
RNA Seq:
Find tissues and cell lines supported by RNA-seq analysis to express GLI2.
Immunogen:
The immunogen is a synthetic peptide directed towards the N terminal region of human GLI2
Species Reactivity:
Human
Predicted Homology Based on Immunogen Sequence:
Human: 100%
Complete computational species homology data:
Anti-GLI2 (ARP31885_T100)
Peptide Sequence:
Synthetic peptide located within the following region: RNDVHLRTPLLKENGDSEAGTEPGGPESTEASSTSQAVEDCLHVRAIKTE
Product Format:
Liquid. Purified antibody supplied in 1x PBS buffer with 0.09% (w/v) sodium azide and 2% sucrose.
Reconstitution and Storage:
For short term use, store at 2-8C up to 1 week. For long term storage, store at -20C in small aliquots to prevent freeze-thaw cycles.
Concentration:
Batch dependent within range: 100 ul at 0.5 - 1 mg/ml
Protein Interactions:
SPOP; SKI; HDAC1; STK36; FBXW11; SUFU; ZIC2; ZIC1; CREB1; TBP;
Blocking Peptide:
For anti-GLI2 (ARP31885_T100) antibody is Catalog # AAP31885 (Previous Catalog # AAPP02680)
Datasheets/Manuals:
Printable datasheet for anti-GLI2 (ARP31885_T100) antibody
Target Reference:
Ikram,M.S., et al., (2004) J. Invest. Dermatol. 122 (6), 1503-1509
Publications:

Kim, Y. et al. Selective down-regulation of glioma-associated oncogene 2 inhibits the proliferation of hepatocellular carcinoma cells. Cancer Res. 67, 3583-93 (2007). IHC, WB, Human 17440069

Kiprilov, E. N. et al. Human embryonic stem cells in culture possess primary cilia with hedgehog signaling machinery. J. Cell Biol. 180, 897-904 (2008). IHC, WB, Human 18332216

Kabasawa, Y. et al. Amelogenin positive cells scattered in the interstitial component of odontogenic fibromas. J. Clin. Pathol. 61, 851-5 (2008). IHC, WB, Human 18344235

Omenetti, A. et al. The hedgehog pathway regulates remodelling responses to biliary obstruction in rats. Gut 57, 1275-82 (2008). IHC, WB, Human 18375471

Omenetti, A. et al. Hedgehog signaling regulates epithelial-mesenchymal transition during biliary fibrosis in rodents and humans. J. Clin. Invest. 118, 3331-42 (2008). IHC, WB, Human 18802480

Takanaga, H. et al. Gli2 is a novel regulator of sox2 expression in telencephalic neuroepithelial cells. Stem Cells 27, 165-74 (2009). IHC, WB, Human 18927476

Madison, B. B., McKenna, L. B., Dolson, D., Epstein, D. J. & Kaestner, K. H. FoxF1 and FoxL1 link hedgehog signaling and the control of epithelial proliferation in the developing stomach and intestine. J. Biol. Chem. 284, 5936-44 (2009). IHC, WB, Human 19049965

Wall, D. S. et al. Progenitor cell proliferation in the retina is dependent on Notch-independent Sonic hedgehog/Hes1 activity. J. Cell Biol. 184, 101-12 (2009). IHC, WB, Human 19124651

Syn, W.-K. et al. Role for hedgehog pathway in regulating growth and function of invariant NKT cells. Eur. J. Immunol. 39, 1879-92 (2009). IHC, WB, Human 19544307

Kim, J. E. et al. Sonic hedgehog signaling proteins and ATP-binding cassette G2 are aberrantly expressed in diffuse large B-cell lymphoma. Mod. Pathol. 22, 1312-20 (2009). IHC, WB, Human 19593328

Ho, L. et al. Gli2 and p53 cooperate to regulate IGFBP-3- mediated chondrocyte apoptosis in the progression from benign to malignant cartilage tumors. Cancer Cell 16, 126-36 (2009). IHC, WB, Human 19647223

Kim, W.-Y. et al. GSK-3 is a master regulator of neural progenitor homeostasis. Nat. Neurosci. 12, 1390-7 (2009). IHC, WB, Human 19801986

Nakamura, K. et al. Hedgehog promotes neovascularization in pancreatic cancers by regulating Ang-1 and IGF-1 expression in bone-marrow derived pro-angiogenic cells. PLoS One 5, e8824 (2010). IHC, WB, Human 20098680

Youssef, K. K. et al. Identification of the cell lineage at the origin of basal cell carcinoma. Nat. Cell Biol. 12, 299-305 (2010). IHC, WB, Human 20154679

Peacock, Z. S., Cox, D. & Schmidt, B. L. Involvement of PTCH1 mutations in the calcifying epithelial odontogenic tumor. Oral Oncol. 46, 387-92 (2010). IHC, WB, Human 20371205

Coon, V. et al. Molecular therapy targeting Sonic hedgehog and hepatocyte growth factor signaling in a mouse model of medulloblastoma. Mol. Cancer Ther. 9, 2627-36 (2010). IHC, WB, Human 20807782

Schneider, S. et al. Expression of the Sonic hedgehog pathway in squamous cell carcinoma of the skin and the mucosa of the head and neck. Head Neck 33, 244-50 (2011). IHC, WB, Human 20848446

Malek, R., Matta, J., Taylor, N., Perry, M. E. & Mendrysa, S. M. The p53 inhibitor MDM2 facilitates Sonic Hedgehog-mediated tumorigenesis and influences cerebellar foliation. PLoS One 6, e17884 (2011). IHC, WB, Human 21437245

Jung, I. H., Jung, D. E., Park, Y. N., Song, S. Y. & Park, S. W. Aberrant Hedgehog ligands induce progressive pancreatic fibrosis by paracrine activation of myofibroblasts and ductular cells in transgenic zebrafish. PLoS One 6, e27941 (2011). IHC, WB, Human 22164219

Yang, L. et al. Hedgehog signaling activation in the development of squamous cell carcinoma and adenocarcinoma of esophagus. Int. J. Biochem. Mol. Biol. 3, 46-57 (2012). IHC, WB, Human 22509480

Tostar, U., Finta, C., Rahman, M. F.-U., Shimokawa, T. & Zaphiropoulos, P. G. Novel mechanism of action on Hedgehog signaling by a suppressor of fused carboxy terminal variant. PLoS One 7, e37761 (2012). IHC, WB, Human 22666390

Bosco-Clément, G. et al. Targeting Gli transcription activation by small molecule suppresses tumor growth. Oncogene 33, 2087-97 (2014). IHC, WB, Human 23686308

Li, Y. et al. Gene expression profiling reveals the heterogeneous transcriptional activity of Oct3/4 and its possible interaction with Gli2 in mouse embryonic stem cells. Genomics 102, 456-67 IHC, WB, Human 24121003

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2 Item(s)

138/05/2018 00:51
  • Overall Experience:
  • Quality:
Human Melanoma tissue in WB

Submitted by:

Antibodies Online Validation No. #101239

Laboratory of Herditary Cancer, Division of Molecular Medicine, Rudjer Boskovic Institute

10/05/17

 

Target: GLI2

Sample Type/Lane Description: Human whole cell lysate

Lane A:  50μg total cell lysate - Melanoma WM793B

Lane B1: 50μg total cell lysate - metastatic Melanoma tissue (endogenously expressed)

Lane B2: 50μg total cell lysate - healthy skin not expressing GLI2

Primary antibody dilution: 1:1000 

Secondary Antibody Dilution: 1:3000

Predicted Molecular Weight: 133kDa

GLI2 FL (full length) Observed Molecular Weight: 250kDa

Isoform GLI2N Observed Molecular Weight: 150kDa

Sample Preparation Methods:

1.       Grow WM793B cells in RPMI supplemented with FBS and penicillin/streptomycin, at 37°C and 5% CO2 in a 10 cm TC-dish.

2.       Wash cells 2x with PBS.

3.       Harvest cells.

4.       Lyse cells in 85µl cold lysis buffer (25mM Tris-HCl pH7.5, 150mM NaCl, 1% NP-40, 1% sodium deoxycholate, 0.1% SDS) with sonication 2x 15s using an Labsonic M sonicator with a 1mm probe 1 cycle, 80% amplitude.

5.       Determine total protein content of the lysates using a BCA protein assay.

 

Protocol:

1.       Denature 50μg of total proteins for 5min at 95°C in Laemmli SDS sample buffer and subsequently separate them on a freshly cast denaturing 7% PAA gel for 2h at 100V.

2.       Transfer proteins onto Amersham Protran Premium 0.2 NC membrane with a Mini Trans-Blot cell for 80min at 100V.

3.       Block the membrane with blocking buffer and blocking reagent (TBST/5% non-fat milk) for 30min at RT.

4.       Incubation with primary rabbit anti human GLI2 antibody (lot QC492960-42551) diluted 1:1000 in TBST/5% non-fat milk ON at 4°C.

5.       Wash membrane 3x for 5min with TBST.

6.       Incubation with secondary anti-rabbit IgG HRP-linked antibody (Cell Signaling Technology, 7074) diluted 1:3000 in 5% non-fat milk/TBST for 1h at RT.

7.       Wash membrane 3x for 5mins with TBST.

8.       Reveal protein bands using Super Signal West Pico and Super Signal West Femto reagents on an UVITEC Alliance Imaging system (Cambridge).

Experimental Notes:

Aviva’s GLI2 antibody (lot QC492960-42551) reveals two protein bands of the expected molecular weight for the full-length (GLI2 FL) and the GLI2 N-terminally truncated (GLI2ΔN) isoform in lysates of WM793B cells.

Aviva’s GLI2 antibody (lot QC492960-42551) also detects both GLI2 FL as well as GLI2ΔN in metastatic melanoma cell lysates whereas neither protein is revealed in human healthy skin tissue in which expression of GLI2 is not to be expected.

The predicted molecular weight of the full length GLI2 is 197kDa which is in accordance with our observation. With our previous anti-GLI2 antibody (sc-20291 (G-20), Santa Cruz Biotechnology) we also detected the GLI2ΔN protein at ~150kDa. The difference in apparent MW could be due to different migration of the marker.

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302/10/2017 07:39
  • Overall Experience:
  • Quality:
Excellent antibody, high specificity!

 

1.            What species + tissue/cell type

Human colon cancer tissue sections

2.            Fixation method

The sections were formalin fixed and paraffin embedded

3.            Antigen retrieval method used

The sections were incubated in citrate buffer, pH 6.0 for 20 minutes at 99-100°C and then cooled at room temperature for 20 minutes

4.            Primary antibody dilution

The GLI2 antibody (ARP31885_T100) was diluted 1:100

5.            Secondary antibody                                 

Sections were incubated for 10 minutes at room temperature with biotinylated anti-rabbit and anti-mouse immunoglobulins, then washed 3 times with 1xTBST and incubated with Streptavidin conjugated to horseradish peroxidase for 10 minutes at room-temperature (Dako, LSAB2 System-HRP)

6.            Secondary antibody dilution

The secondary antibody from the kit is ready-to-use.

7.            Description of Stains and counterstain

The sections were stained with DAB chromogen (Dako, Liquid DAB+ Substrate Chromogen System) and counterstained with haematoxylin (Biognost).

 

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