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Antibody Customer Review for GLI1 Polyclonal Antibody (STJ110685)

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Originally isolated in glioblastoma, Gli1 binds to the DNA consensus sequence 5'-GACCACCCA-3'. The Gli proteins are effectors of Hedgehog (Hh) signalling, and play a role in cell fate determination. proliferation and organ cell patterning during embryo development. In the developing spinal cord the target genes of Gli proteins, that are themselves transcription factors, are arranged into a complex gene regulatory network that translates the extracellular concentration gradient of Sonic hedgehog into different cell fates along the dorsoventral axis.

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Antibody Customer Review for GLI1 Polyclonal Antibody (STJ110685)

  1. 1. HOW DID OUR ANTIBODY PERFORM? ANTIBODY CUSTOMER REVIEW: Gli1 Polyclonal Antibody (STJ110685)
  2. 2. WHAT WAS USED? Primary antibody: Gli1 Polyclonal Antibody (STJ110685) Provider: St John's Laboratory Dilution ratio: 1:200 Application: Immunohistochemistry Materials for validation: Interganglionic region (MGE/LGE) E12.5 ventral telencephalon
  3. 3. WHAT WAS THE PROTOCOL? Treatment of materials: Tissue was fixed with 4% PFA in PBS for 2.5 hours at 4C, then incubated in 20% sucrose/PBS at 4C until tissue sunk (~2 hours). Thereafter, the tissue was embedded in cryomold solution (OCT) and flash frozen using cold isopentanol (cool with dry ice) Wash: 1X wash for 5 minutes with 0.1% Triton X-100 PBS. Blocking: 1 hour blocking with a solution containing 10% Donkey Serum, 1% BSA, 1M glycine, in 0.5% Triton X-100 PBS. Primary Ab Application: 48 hr incubation with primary antibody at 4C in a solution containing 1% BSA in 0.1% Triton X-100 PBS. Wash: 4X washes, 5 minutes each, with 0.1% Triton X-100 PBS. Secondary Ab Application: 1 hour incubation with 2nd antibody Alexa 488 (Invitrogen), 1:1500 dilution in 0.1% Triton X-100 PBS. Wash: 3X washes, 5 minutes each, with 0.1% Triton X-100 PBS.
  4. 4. WHAT DID THE CUSTOMER THINK? Testing results provided by A. Molero of Albert Einstein College of Medicine said “low nuclear signal but high cytoplasmic signal. Antigen retrieval with heat/citrate buffer increase the cytoplasmic signal but notably reduces nuclear immunoreactivity.”
  5. 5. JOIN THE ANTIBODY VALIDATION PROJECT
  6. 6. 1. Order any STJ9 catalogued product using this online form. 2. Test it in your laboratory using your preferred method. 3. Return a fully completed review form to receive rewards. …in 3 simple steps…
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