GLI-3 Antibody - BSA Free

Novus Biologicals | Catalog # NBP2-29627

Novus Biologicals
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Key Product Details

Validated by

Biological Validation

Species Reactivity

Human, Mouse

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Flow Cytometry, Flow (Intracellular), Immunocytochemistry/ Immunofluorescence

Cited:

Immunohistochemistry-Paraffin

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all GLI-3 Primary Antibodies »

Product Specifications

Immunogen

A synthetic peptide made to an internal portion of human Gli3 protein (between residues 50-120) [UniProt P10071]

Reactivity Notes

Immunogen displays the following percentage of sequence identity for non-tested species: Xenopus (84%) and Chicken (85%).

Localization

Nucleus. Cytoplasm.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

190 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for GLI-3 Antibody - BSA Free

Western Blot: GLI-3 AntibodyBSA Free [NBP2-29627]

Western Blot: GLI-3 AntibodyBSA Free [NBP2-29627]

Western Blot: GLI-3 Antibody [NBP2-29627] - Adult mouse brain tissue lysates. Image from verified customer review.
Immunocytochemistry/ Immunofluorescence: GLI-3 Antibody - BSA Free [NBP2-29627]

Immunocytochemistry/ Immunofluorescence: GLI-3 Antibody - BSA Free [NBP2-29627]

Immunocytochemistry/Immunofluorescence: GLI-3 Antibody [NBP2-29627] - HeLa cells with FITC (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and DyLight 550 (red).
Immunohistochemistry-Paraffin: GLI-3 Antibody - BSA Free [NBP2-29627]

Immunohistochemistry-Paraffin: GLI-3 Antibody - BSA Free [NBP2-29627]

Immunohistochemistry-Paraffin: GLI-3 Antibody [NBP2-29627] - Analysis of in mouse liver.
Flow (Intracellular): GLI-3 Antibody - BSA Free [NBP2-29627]

Flow (Intracellular): GLI-3 Antibody - BSA Free [NBP2-29627]

Flow (Intracellular): GLI-3 Antibody [NBP2-29627] - Intracellular staining for GLI-3 in untreated human B cell lines (BCWM.1, MWCL-1 and RPCI-WM1) using anti-GLI-3 antibody. Rabbit IgG Isotype Control (Cat# NBP2-36463) was used as a negative control. Image courtesy of Sherine Elsawa, Northern Illinois University.
Western Blot: GLI-3 AntibodyBSA Free [NBP2-29627]

Western Blot: GLI-3 AntibodyBSA Free [NBP2-29627]

Western Blot: GLI-3 Antibody [NBP2-29627] - Analysis of GLI-3 in Hela cell lysate and MEF cell lysate.
Immunohistochemistry-Paraffin: GLI-3 Antibody - BSA Free [NBP2-29627]

Immunohistochemistry-Paraffin: GLI-3 Antibody - BSA Free [NBP2-29627]

Immunohistochemistry-Paraffin: GLI-3 Antibody [NBP2-29627] - Analysis of in mouse testis.

Applications for GLI-3 Antibody - BSA Free

Application
Recommended Usage

Flow Cytometry

reported in scientific literature (PMID 26238488)

Immunocytochemistry/ Immunofluorescence

1:50

Immunohistochemistry

1:100

Immunohistochemistry-Paraffin

1:100

Western Blot

1:1000
Application Notes
In Western blot, bands were observed at ~75 kDa (GLI3R) in both HeLa and MEF cell lysates while an additional band was observed ~150 kDa (GLI3A) in MEF cell lysate only. The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Reviewed Applications

Read 1 review rated 4 using NBP2-29627 in the following applications:

Flow Cytometry Panel Builder

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Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

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-3

Gli-3 (also known as Zinc Finger Protein Gli-3 or GLI-Kruppel family member GLI-3) belongs to the GLI C2H2-type zinc-finger protein family and contains 5 C2H2-type zinc fingers. Gli-3 is very important for normal limb and brain development and is implicated in the transduction of Shh signal. Gli-3 is a nuclear protein expressed in a wide variety of normal adult tissues, including lung, colon, spleen, placenta, testis, and myometrium. Defects in Gli-3 are the cause of Greig cephalo-polysyndactyly syndrome (GCPS); an autosomal dominant disorder-affecting limb and craniofacial development. Two isoforms of human Gli-3 have been reported. One is the full length protein at ~170-190kDa and the other is a truncated isoform at ~80kDa.

Long Name

GLI-Kruppel family member GLI3

Alternate Names

ACLS, ADD, Bph, GCPS, GLI3, PAPA1, PAPB, Pdn, PHS, PPDIV, Xt

Entrez Gene IDs

2737 (Human)

Gene Symbol

GLI3

UniProt

P10071

Additional GLI-3 Products

Product Documents for GLI-3 Antibody - BSA Free

Certificate of Analysis

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Product Specific Notices for GLI-3 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.

Citations for GLI-3 Antibody - BSA Free

Customer Reviews for GLI-3 Antibody - BSA Free (1)

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  • GLI-3 Antibody
    Name: Zhihui Zhu
    Application: Western Blot
    Sample Tested: mouse brain lysates and Adult mouse brain tissue
    Species: Mouse
    Verified Customer | Posted 07/09/2019
    Azure c600
    mice brain samples for western blot.
    GLI-3 Antibody - BSA Free NBP2-29627

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Protocols

View specific protocols for GLI-3 Antibody - BSA Free (NBP2-29627):

GLI-3 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.

GLI-3 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 4 C.
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.


GLI-3 Antibody:
Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 25 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 anti-Gli3 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 anti-rabbit 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%.

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Associated Pathways

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