Analysis Note

ControlIncludes negative control rabbit IgG and control primers specific for human dihydrofolate reductase (DHFR) promoter.

Application

Western Blot Analysis:
HeLa nuclear extract was resolved by electrophoresis, transferred to nitrocellulose and probed with anti-Sp1 (0.2 µg/mL). Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system (Please see figures).

Research CategoryEpigenetics & Nuclear Function

Research Sub CategoryChromatin Biology

SP1 ChIP validated antibody & primer set including the ChIP-grade antibody & the specific control PCR primers used for chromatin immunoprecipitation of SP1.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

Transcription factor SP1 can activate or repress transcription in response to physiological and pathological stimuli. Binds with high affinity to GC-rich motifs and regulates the expression of a large number of genes involved in a variety of processes such as cell growth, apoptosis, differentiation and immune responses. Highly regulated by post-translational modifications (phosphorylations, sumoylation, proteolytic cleavage, glycosylation and acetylation). Binds also the PDGFR-alpha G-box promoter. May have a role in modulating the cellular response to DNA damage. Implicated in chromatin remodeling. Plays a role in the recruitment of SMARCA4/BRG1 on the c-FOS promoter. Plays an essential role in the regulation of FE65 gene expression. In complex with ATF7IP, maintains telomerase activity in cancer cells by inducing TERT and TERC gene expression. Isoform 3 is a stronger activator of transcription than isoform 1. Positively regulates the transcription of the core clock component ARNTL/BMAL1.

Every lot of the ChIPAb+ line of antibodies is individually validated for chromatin precipitation, in order to guarantee successful ChIP assays every time. Each antibody includes a control primer set for performance confirmation. Sp1 antibody and the negative control antibody (rabbit normal IgG) can be used to demonstrate that the Sp1 antibody is functionally validated in the precipitation of Sp1 associated chromatin.
The qPCR primers included flank the Sp1 binding site in human DHFR promoter.

Immunogen

Full length human Sp1 protein

Legal Information

UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

Packaging

25 assays per kit, ~4µg per chromatin immunoprecipitation.

Physical form

Protein A purified

Anti-Sp1 (Rabbit polyclonal IgG). 1 vial containing 100 µg of protein A purified IgG in 100 µL of 0.014 M phosphate buffer, pH 7.6, 0.175 M NaCl, 0.07% sodium azide and 30% glycerol. Store at -20°C. The Sp1 antibody is made against full length human Sp1 protein and can recognize Sp1, MW 105 kDa and 95 kDa, of human, mouse and rat origins.

Normal Rabbit IgG. One vial containing 125 µg of normal rabbit IgG in 125 µL. Store at -20°C.

ChIP primers DHFR. 1 vial containing 75 µL of 5 µM of each control primer specific for human dihydrofolate reductase (DHFR) promoter. Store at -20°C.
FOR: TCG CCT GCA CAA ATA GGG AC
REV: AGA ACG CGC GGT CAA GTT T

Format: Purified

Quality

Chromatin immunoprecipitation:
Sonicated chromatin prepared from 1 x 10⁶ HeLa cells was subjected to chromatin immunoprecipitation using 4 µg of either the negative control antibody, Rabbit Normal IgG or Rabbit Anti-Sp1 and the Magna ChIPTM A kit (Cat.# 17-610) Rapid Protocol (2 hour IP). Successful enrichment of Sp1 associated DNA fragments was verified by qPCR using ChIP Primers DHFR flanking the human DHFR promoter that contains a Sp1 binding site (Please see figures).
Please refer to the EZ-Magna ChIP™ A (Cat.# 17-408)
or EZ-ChIP™ (Cat.# 17-371) kit protocols for
experimental details.

Specificity

Sp1

Storage and Stability

1 year at -20°C from date of shipment.

Target description

105 kDa and 95 kDa