Antibody Customer Review for SP1 Polyclonal Antibody (STJ95738)


Target information

SP1 is a Zinc-finger transcription factor which binds with high affinity to GC-rich motifs to regulate genes which are involved in various processes like apoptosis, differentiation, growth and immune response. This protein is regulated by post translational modifications such as phosphorylation, sumoylation, proteolytic cleavage, glycosylation and acetylation. It is thought to play a role in the cellular response to DNA damage and implicated in chromatin remodeling.

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1. HOW DID OUR ANTIBODY PERFORM? ANTIBODY CUSTOMER REVIEW: SP1 Polyclonal Antibody (STJ95738)
2. WHAT WAS USED? Primary antibody: SP1 PolyclonalAntibody (STJ95738) Provider: St John’s Laboratory Dilution ratio: 1:3000 Application: Immunohistochemistry Materials for validation: E12.5 telencephalon
3. WHAT WAS THE PROTOCOL? Does not require antigen retrieval. Pre-treatment with heat citrate or EDTA buffer antigen retrieval reduces the signal to noise ratio. 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. WHAT DID THE CUSTOMER THINK? Testing results provided by A. Molero of The Saul Korey Department of Neurology said that there was ¡°excellent signal to noise ratio, low background¡±.
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