Phospho-Gli1 (Thr1074) Antibody | Affinity Biosciences

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Western blot analysis of extracts from various samples, using Phospho-Gli1 (Thr1074) Antibody.

$Figure 6 PRKCI mediated radiosensitivity in CC via the Hh signaling pathway to change GLI1 localization. (A) The viability of the si-GLI1-transfected HeLa-ov-PRKCI and SiHa-ov-PRKCI cells after radiotherapy at 8 Gy determined by CCK-8 assays. (B) Western blotting assays were used to assess the protein expression of PRKCI when GLI1 expression was downregulated by siRNA in both the HeLa-ov-PRKCI and SiHa-ov-PRKCI cells. (C) The relative protein localization of GLI1 after transfection with shRNA-con and shRNA-PRKCI or ov-con and ov-PRKCI in the HeLa and SiHa cells treated with radiation was determined by immunofluorescence assays. (D) Western blotting analysis of GLI1 and p-GLI1 protein levels in the shRNA-PRKCI- and ov-PRKCI-transfected HeLa and SiHa cells. (E) Western blotting analysis of GLI1 and p-GLI1 protein levels in the cytoplasmic and nuclear fractions of the shRNA-PRKCI- and ov-PRKCI-transfected HeLa and SiHa cells. Scale bars are shown. Data are shown as the mean ± SD from three independent experiments. NS, not significant. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001 by multiple t-tests (A). CC, cervical cancer; CCK-8, Cell Counting Kit-8.$

Figure 6 PRKCI mediated radiosensitivity in CC via the Hh signaling pathway to change GLI1 localization.

Product:	Phospho-Gli1 (Thr1074) Antibody
Catalog:	AF7070
Description:	Rabbit polyclonal antibody to Phospho-Gli1 (Thr1074)
Application:	WB
Reactivity:	Human, Mouse
Prediction:	Pig, Zebrafish, Bovine, Horse, Sheep, Rabbit, Dog, Chicken
Mol.Wt.:	119kDa; 118kD(Calculated).
Uniprot:	P08151
RRID:	AB_2843510

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Blocking peptides are peptides that bind specifically to the target antibody and block antibody binding. These peptide usually contains the epitope recognized by the antibody. Antibodies bound to the blocking peptide no longer bind to the epitope on the target protein. By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific.

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Data Sheet

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IHC Protocol

IF/ICC Protocol

Product Info

Source:

Rabbit

Application:

WB 1:500-1:2000
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:

Human,Mouse

Prediction:

Pig(90%), Zebrafish(80%), Bovine(90%), Horse(90%), Sheep(90%), Rabbit(80%), Dog(90%), Chicken(100%)

Clonality:

Polyclonal

Specificity:

Phospho-Gli1 (Thr1074) Antibody detects endogenous levels of Gli1 only when phosphorylated at Thr1074.

RRID:

AB_2843510
Cite Format: Affinity Biosciences Cat# AF7070, RRID:AB_2843510.

Conjugate:

Unconjugated.

Purification:

The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.

Storage:

Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.

Alias:

Fold/Unfold

Gli 1; GLI; GLI family zinc finger 1; GLI Kruppel family member 1; gli1; GLI1_HUMAN; Glioma associated oncogene 1; Glioma associated oncogene homolog 1 (zinc finger protein); Glioma associated oncogene homolog; Glioma-associated oncogene; Oncogene GLI; Zfp 5; Zfp5; Zinc finger protein GLI 1; Zinc finger protein GLI1;

Immunogens

Immunogen:

Uniprot:

P08151(GLI1_HUMAN) >>Visit HPA database.

Gene(ID):

GLI1(2735)

Expression:

P08151 GLI1_HUMAN:

Detected in testis (at protein level) (PubMed:2105456). Testis, myometrium and fallopian tube. Also expressed in the brain with highest expression in the cerebellum, optic nerve and olfactory tract (PubMed:19878745). Isoform 1 is detected in brain, spleen, pancreas, liver, kidney and placenta; isoform 2 is not detectable in these tissues (PubMed:19706761).

Sequence:

P08151 GLI1_HUMAN:
Protein BLAST With
NCBI/
ExPASy/
Uniprot

MFNSMTPPPISSYGEPCCLRPLPSQGAPSVGTEGLSGPPFCHQANLMSGPHSYGPARETNSCTEGPLFSSPRSAVKLTKKRALSISPLSDASLDLQTVIRTSPSSLVAFINSRCTSPGGSYGHLSIGTMSPSLGFPAQMNHQKGPSPSFGVQPCGPHDSARGGMIPHPQSRGPFPTCQLKSELDMLVGKCREEPLEGDMSSPNSTGIQDPLLGMLDGREDLEREEKREPESVYETDCRWDGCSQEFDSQEQLVHHINSEHIHGERKEFVCHWGGCSRELRPFKAQYMLVVHMRRHTGEKPHKCTFEGCRKSYSRLENLKTHLRSHTGEKPYMCEHEGCSKAFSNASDRAKHQNRTHSNEKPYVCKLPGCTKRYTDPSSLRKHVKTVHGPDAHVTKRHRGDGPLPRAPSISTVEPKREREGGPIREESRLTVPEGAMKPQPSPGAQSSCSSDHSPAGSAANTDSGVEMTGNAGGSTEDLSSLDEGPCIAGTGLSTLRRLENLRLDQLHQLRPIGTRGLKLPSLSHTGTTVSRRVGPPVSLERRSSSSSSISSAYTVSRRSSLASPFPPGSPPENGASSLPGLMPAQHYLLRARYASARGGGTSPTAASSLDRIGGLPMPPWRSRAEYPGYNPNAGVTRRASDPAQAADRPAPARVQRFKSLGCVHTPPTVAGGGQNFDPYLPTSVYSPQPPSITENAAMDARGLQEEPEVGTSMVGSGLNPYMDFPPTDTLGYGGPEGAAAEPYGARGPGSLPLGPGPPTNYGPNPCPQQASYPDPTQETWGEFPSHSGLYPGPKALGGTYSQCPRLEHYGQVQVKPEQGCPVGSDSTGLAPCLNAHPSEGPPHPQPLFSHYPQPSPPQYLQSGPYTQPPPDYLPSEPRPCLDFDSPTHSTGQLKAQLVCNYVQSQQELLWEGGGREDAPAQEPSYQSPKFLGGSQVSPSRAKAPVNTYGPGFGPNLPNHKSGSYPTPSPCHENFVVGANRASHRAAAPPRLLPPLPTCYGPLKVGGTNPSCGHPEVGRLGGGPALYPPPEGQVCNPLDSLDLDNTQLDFVAILDEPQGLSPPPSHDQRGSSGHTPPPSGPPNMAVGNMSVLLRSLPGETEFLNSSA

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species

Results

Score

Chicken

100

Pig

Horse

Bovine

Sheep

Dog

Zebrafish

Rabbit

Xenopus

Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P08151 As Substrate

P08151

Site	PTM Type	Enzyme	Source
	Phosphorylation		Uniprot
	Ubiquitination		Uniprot
T63	Phosphorylation		Uniprot
S69	Phosphorylation		Uniprot
S84	Phosphorylation	P23443 (RPS6KB1)	Uniprot
S102	Phosphorylation	P54646 (PRKAA2)	Uniprot
S130	Phosphorylation		Uniprot
R161	Methylation		Uniprot
S201	Phosphorylation		Uniprot
S204	Phosphorylation		Uniprot
T205	Phosphorylation		Uniprot
S243	Phosphorylation	P41743 (PRKCI)	Uniprot
T304	Phosphorylation	P41743 (PRKCI)	Uniprot
K319	Ubiquitination		Uniprot
K365	Acetylation		Uniprot
K371	Acetylation		Uniprot
T374	Phosphorylation	P17612 (PRKACA)	Uniprot
S408	Phosphorylation	P54646 (PRKAA2)	Uniprot
K518	Acetylation		Uniprot
S521	Phosphorylation		Uniprot
T527	Phosphorylation		Uniprot
S538	Phosphorylation		Uniprot
S544	Phosphorylation	P17612 (PRKACA)	Uniprot
S545	Phosphorylation		Uniprot
S560	Phosphorylation	P17612 (PRKACA)	Uniprot
R597	Methylation		Uniprot
S602	Phosphorylation		Uniprot
S622	Phosphorylation		Uniprot
S640	Phosphorylation	P17612 (PRKACA)	Uniprot
S934	Phosphorylation		Uniprot
S937	Phosphorylation		Uniprot
S939	Phosphorylation		Uniprot
S968	Phosphorylation		Uniprot
S1071	Phosphorylation		Uniprot
T1074	Phosphorylation	P54646 (PRKAA2)	Uniprot
S1078	Phosphorylation		Uniprot

Research Backgrounds

P08151_GLI1_HUMAN

Function:

Acts as a transcriptional activator. Binds to the DNA consensus sequence 5'-GACCACCCA-3'. Regulates the transcription of specific genes during normal development. Plays a role in craniofacial development and digital development, as well as development of the central nervous system and gastrointestinal tract. Mediates SHH signaling. Plays a role in cell proliferation and differentiation via its role in SHH signaling.

Acts as a transcriptional activator, but activates a different set of genes than isoform 1. Activates expression of CD24, unlike isoform 1. Mediates SHH signaling. Promotes cancer cell migration.

PTMs:

Phosphorylated in vitro by ULK3.

Acetylation at Lys-518 down-regulates transcriptional activity. Deacetylated by HDAC1.

Subcellular Location:

Cytoplasm. Nucleus.
Note: Tethered in the cytoplasm by binding to SUFU (PubMed:10806483). Activation and translocation to the nucleus is promoted by interaction with STK36 (PubMed:10806483). Phosphorylation by ULK3 may promote nuclear localization (PubMed:19878745). Translocation to the nucleus is promoted by interaction with ZIC1 (PubMed:11238441).

Cytoplasm. Nucleus.

Tissue Specificity:

Detected in testis (at protein level). Testis, myometrium and fallopian tube. Also expressed in the brain with highest expression in the cerebellum, optic nerve and olfactory tract. Isoform 1 is detected in brain, spleen, pancreas, liver, kidney and placenta; isoform 2 is not detectable in these tissues.

Subunit Structure:

Interacts with KIF7 (By similarity). Interacts with STK36. Interacts with ZIC1; the interaction enhances transcription activation. Interacts with SUFU; this inhibits transcriptional activation by GLI1.

Family&Domains:

Belongs to the GLI C2H2-type zinc-finger protein family.

Research Fields

· Environmental Information Processing > Signal transduction > cAMP signaling pathway. (View pathway)

· Environmental Information Processing > Signal transduction > Hedgehog signaling pathway. (View pathway)

· Human Diseases > Cancers: Overview > Pathways in cancer. (View pathway)

· Human Diseases > Cancers: Specific types > Basal cell carcinoma. (View pathway)

References

1). PRKCI Mediates Radiosensitivity via the Hedgehog/GLI1 Pathway in Cervical Cancer. Frontiers in Oncology, 2023 (PubMed: 35785194) [IF=4.7]

Application: WB Species: Human Sample: HeLa cells

Figure 6 PRKCI mediated radiosensitivity in CC via the Hh signaling pathway to change GLI1 localization. (A) The viability of the si-GLI1-transfected HeLa-ov-PRKCI and SiHa-ov-PRKCI cells after radiotherapy at 8 Gy determined by CCK-8 assays. (B) Western blotting assays were used to assess the protein expression of PRKCI when GLI1 expression was downregulated by siRNA in both the HeLa-ov-PRKCI and SiHa-ov-PRKCI cells. (C) The relative protein localization of GLI1 after transfection with shRNA-con and shRNA-PRKCI or ov-con and ov-PRKCI in the HeLa and SiHa cells treated with radiation was determined by immunofluorescence assays. (D) Western blotting analysis of GLI1 and p-GLI1 protein levels in the shRNA-PRKCI- and ov-PRKCI-transfected HeLa and SiHa cells. (E) Western blotting analysis of GLI1 and p-GLI1 protein levels in the cytoplasmic and nuclear fractions of the shRNA-PRKCI- and ov-PRKCI-transfected HeLa and SiHa cells. Scale bars are shown. Data are shown as the mean ± SD from three independent experiments. NS, not significant. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001 by multiple t-tests (A). CC, cervical cancer; CCK-8, Cell Counting Kit-8.

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Phospho-Gli1 (Thr1074) Antibody - #AF7070