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Synthetic peptide (Unfortunately the amino acid sequence is considered to be commercially sensitive). The peptide sequence is within the region between aa750 and the C-terminus (aa785) of human SP1 using the numbering given in entry XP_028606.4 (GeneID 666).
Our Abpromise guarantee covers the use of ab13370 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IP||Use at 1-4 µg/mg of lysate.|
|WB||1/2500 - 1/10000. Detects a band of approximately 90 kDa (predicted molecular weight: 90 kDa).|
|IHC-P||Use at an assay dependent concentration.|
ChIP analysis using ab13370 binding SP1 in Human primary macrophage. Cells were cross-linked for 15 minutes with formaldehyde. Samples were incubated with primary antibody (1/500) for 12 hours at 4°C. Protein binding was detected using real-time PCR.
SP1 was immunoprecipitated from HeLa cells whole cell lysate with ab13370. Anti-SP1 antibody - ChIP Grade (ab13370) used at 6µg per reaction. SP1 was also immunoprecipitated by a previous lot of this antibody. For blotting immunoprecipitated SP1, ab13370 was used at 1µg/ml. Chemiluminescence with an exposure time of 10 seconds.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue sections labeling SP1 with ab13370 at 1/5000 dilution. DAB detection.
Detection of Human SP1 by Western Blot and Immunoprecipitation. Samples: A. Whole cell lysate (50 mcg for E; 10 mcg for T) from HEK 293T cells that were mock transfected (E) or transfected with a Sp1 expression construct (T). B. Whole cell lysate (700 mcg) from HEK 293T cells. Antibody: A. Affinity purified rabbit anti-Sp1 antibody (ab13370) used at the indicated concentrations. B. SP1 was immunoprecipitated using affinity purified rabbit anti-SP1 antibodies (lane 2: ab13405, lane 3: ab13370; lane 1: SP-1 antibody, details unknown) using each antibody at 3.75 mcg/mg lysate. Subsequently, ab13370 was used at 0.04 mcg/ml for WB. Detection: Chemiluminescence with an exposure time of 10 seconds (A) or 1 min (B).