AF3526

antibody from R&D Systems

Targeting: GLI2 HPE9, THP1, THP2

Western blot (Data presented on provider website)

Immunohistochemistry (Data presented on provider website)

Chromatin Immunoprecipitation (Supportive data in Antibodypedia)

Antibody Data

Product number

AF3526

Provider

R&D Systems

Product name

Human GLI-2 Antibody

Antibody type

Polyclonal

Description

Immunogen affinity purified. Detects human GLI-2 in direct ELISAs and Western blots. In direct ELISAs and Western blots, approximately 25% cross-reactivity with recombinant mouse GLI-2 and 5% cross-reactivity with recombinant human (rh) GLI-1 and rhGLI-3 is observed.

Reactivity

Human

Host

Sheep

Conjugate

Unconjugated

Antigen sequence

BAA25668


Isotype

IgG

Vial size

100 ug

Concentration

LYOPH

Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles. 12 months from date of receipt, -20 to -70 °C as supplied. 1 month, 2 to 8 °C under sterile conditions after reconstitution. 6 months, -20 to -70 °C under sterile conditions after reconstitution.

References

Submitted references

Regulation of GLI1 by cis DNA elements and epigenetic marks.

Taylor R, Long J, Yoon JW, Childs R, Sylvestersen KB, Nielsen ML, Leong KF, Iannaccone S, Walterhouse DO, Robbins DJ, Iannaccone P
DNA repair 2019 Jul;79:10-21

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SOX2 regulates self-renewal and tumorigenicity of human melanoma-initiating cells.

Santini R, Pietrobono S, Pandolfi S, Montagnani V, D'Amico M, Penachioni JY, Vinci MC, Borgognoni L, Stecca B
Oncogene 2014 Sep 18;33(38):4697-708

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GLI2 transcription factor mediates cytokine cross-talk in the tumor microenvironment.

Elsawa SF, Almada LL, Ziesmer SC, Novak AJ, Witzig TE, Ansell SM, Fernandez-Zapico ME
The Journal of biological chemistry 2011 Jun 17;286(24):21524-34

Chromatin Immunoprecipitation

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

Detection of GLI-2-regulated Genes by Chromatin Immunoprecipitation. Jurkat human acute T cell leukemia cell line treated with 50 ng/mL PMA and 200 ng/mL calcium ionomycin for 30 minutes was fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. GLI-2/DNA complexes were immunoprecipitated using 5 μg Sheep Anti-Human GLI-2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3526) or control antibody (Catalog # 5-001-A) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Sheep IgG Secondary Antibody (Catalog # BAF016). Immunocomplexes were captured using 50 μL of MagCellect Streptavidin Ferrofluid (Catalog # MAG999) and DNA was purified using chelating resin solution. The bcl-2 promoter was detected by standard PCR.