Skip to main content

Key Product Details

Validated by

Biological Validation

Species Reactivity

Human, Mouse

Applications

Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, In-situ Hybridization, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free

Concentration

1.0 mg/ml

Product Summary for GLI-1 Antibody - BSA Free

Immunogen

A synthetic peptide made to an internal portion of the human Gli1 protein (between residues 150-200) [UniProt P08151]

Localization

Cytoplasm. Nucleus. Note: Tethered in the cytoplasm by binding to SUFU. Activation and translocation to the nucleus is promoted by interaction with STK36. Phosphorylation by ULK3 may promote nuclear localization.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for GLI-1 Antibody - BSA Free

Immunohistochemistry: GLI-1 Antibody [NBP1-78259] - In vivo GLI1 expression after intratumoral administration of NVP-LDE225. LOX OMVI human melanoma cells were injected s.c into both flanks. Tumors were treated intratumorally on daily basis with vehicle (A & B) or NVP-LDE225 (C & D). Immunofluorescent microscopy of GLI1 was performed on isolated tumor tissues. GLI1 staining was performed by overnight incubation of sections at 4C with rabbit anti-GLI-1 polyclonal Ab or isotype control (A & C) followed by an 1 hr-incubation with AF488 Donkey IgG, anti-rabbit at RT (green). Counterstaining of nuclei was performed with propidium iodide (red). Pictures were taken on a confocal laser-scanning microscope system (LSM 410; Zeiss). Yellow color corresponds to double positive (anti-GLI1 and propidium iodide) nuclear staining. Image collected and cropped by CiteAb from the following publication (dx.plos.org/10.1371/journal.pone.0069064) licensed under a CC-BY license.
Western Blot: GLI-1 Antibody [NBP1-78259] - Analysis of GLI-1 on partial recombinant GLI-1 protein (molecular weight of partial recombinant protein is 37 kDa).
Immunocytochemistry/Immunofluorescence: GLI-1 Antibody [NBP1-78259] - GLI1 antibody was tested in HepG2 cells with FITC (green). Nuclei and actin were counterstained with DAPI (blue) and Phalloidin (red).

Applications for GLI-1 Antibody - BSA Free

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:200

Immunohistochemistry

1:200

Immunohistochemistry-Frozen

reported in scientific literature (PMID 25799059)

Immunohistochemistry-Paraffin

1:400

In-situ Hybridization

reported in scientific literature (PMID 24506883)

Western Blot

1:1000
Application Notes
Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (pH 6.0) is recommended.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Published Applications

Read 19 publications using NBP1-78259 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS and 30% Glycerol

Format

BSA Free

Preservative

0.05% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: GLI-1

GLI-1 is a transcription factor involved in development, patterning and the regulation of cell fate-determination. A down-stream effector of the Hedgehog (HH) signaling pathway, GLI proteins work with other transcriptional activators to up-regulate the expression of GLI-response genes. In addition to its role in embryogenesis, the aberrant activation of the HH/GLI pathway has been identified in numerous forms of cancer, implicating the GLI family of proteins as potential therapeutic targets. In the absence of a stimulus, GLI-1 is anchored to the cytoplasm via Cos-2, which is then released and translocates to the nucleus upon activation of the HH pathway. In the nucleus, GLI-1 binds DNA via a highly conserved C2-H2 zinc finger domain and acts exclusively as an activator increasing transcription of GLI1, PTCH1, PTCH2, HHIP1, MYCN, CCND1, CCND2, BCL2, CFLAR, FOXF1, FOXL1, PRDM1 (BLIMP1), JAG2, GREM1, and Follistatin.

Long Name

Glioma-Associated Oncogene Homolog 1 [Zinc Finger Protein]

Alternate Names

GLI1

Entrez Gene IDs

2735 (Human)

Gene Symbol

GLI1

UniProt

Product Documents for GLI-1 Antibody - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for GLI-1 Antibody - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Loading...

Protocols

View specific protocols for GLI-1 Antibody - BSA Free (NBP1-78259):

[[URL:https://www.novusbio.com/products/gli-1-antibody_nbp1-78259]][[Caption:… Antibody]]
Immunocytochemistry Protocol

Culture cells to appropriate density in 35 mm culture dishes or 6-well plates.
1. Remove culture medium and add 10% formalin to the dish. Fix at room temperature for 30 minutes.
2. Remove the formalin and add ice cold methanol. Incubate for 5-10 minutes.
3. Remove methanol and add washing solution (i.e. PBS). Be sure to not let the specimen dry out. Wash three times for 10 minutes.
4. To block nonspecific antibody binding incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
5. Add primary antibody at appropriate dilution and incubate at room temperature from 2 hours to overnight at room temperature.
6. Remove primary antibody and replace with washing solution. Wash three times for 10 minutes.
7. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
8. Remove antibody and replace with wash solution, then wash for 10 minutes. Add Hoechst 33258 to wash solution at 1:25,0000 and incubate for 10 minutes. Wash a third time for 10 minutes.
9. Cells can be viewed directly after washing. The plates can also be stored in PBS containing Azide covered in Parafilm (TM). Cells can also be cover-slipped using Fluoromount, with appropriate sealing.

*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.
[[URL:https://www.novusbio.com/products/gli-1-antibody_nbp1-78259]][[Caption:… Antibody]]
Immunohistochemistry-Paraffin Embedded Sections

Antigen 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:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in wash buffer for 5 minutes.
3. Block each section with 100-400 ul blocking solution for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul biotinylated diluted secondary antibody. Incubate 30 minutes at room temperature.
7. Remove secondary antibody solution and wash sections three times with wash buffer for 5 minutes each.
8. Add 100-400 ul Streptavidin-HRP reagent to each section and incubate for 30 minutes at room temperature.
9. Wash sections three times in wash buffer for 5 minutes each.
10. Add 100-400 ul DAB substrate to each section and monitor staining closely.
11. As soon as the sections develop, immerse slides in deionized water.
12. Counterstain sections in hematoxylin.
13. Wash sections in deionized water two times for 5 minutes each.
14. Dehydrate sections.
15. Mount coverslips.
*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.
[[URL:https://www.novusbio.com/products/gli-1-antibody_nbp1-78259]][[Caption:… Antibody]]
Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 40 ug of total protein per lane.
2. Transfer proteins to membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain according to standard Ponceau S procedure (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot.
5. Block the membrane using standard blocking buffer for at least 1 hour.
6. Wash the membrane in wash buffer three times for 10 minutes each.
7. Dilute primary antibody in blocking buffer and incubate 1 hour at room temperature.
8. Wash the membrane in wash buffer three times for 10 minutes each.
9. Apply the diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.
10. Wash the blot in wash buffer three times for 10 minutes each (this step can be repeated as required to reduce background).
11. 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%.

*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.

FAQs for GLI-1 Antibody - BSA Free

Showing  1 - 1 of 1 FAQ Showing All
  • Q: What protocols did you use to stain mouse tissues (IHC-P) with NBP1-78259? For antigen retrieval, did you use boiling in citric buffer? What did you use for blocking? What do you dilute the antibodies in and what is the dilution when you are incubating with NBP1-78259?

    A: Please see this link to our specific protocol for using this antibody in IHC. I think this should provide you with the answers you needed.

Showing  1 - 1 of 1 FAQ Showing All
Loading...