WB result of SP1 Recombinant Rabbit mAb
Primary antibody: SP1 Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: HeLa whole cell lysate 20 µg
Lane 2: 293T whole cell lysate 20 µg
Lane 3: Ramos whole cell lysate 20 µg
Secondary antibody: Goat Anti- rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 81 kDa
Observed MW: 100 kDa
SP1 Recombinant Rabbit mAb (S-4102)
SP1 Recombinant Rabbit mAb (S-4102)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | SP1 |
| Synonyms | Transcription factor Sp1; TSFP1 |
| Location | Cytoplasm, Nucleus |
| Accession | P08047 |
| Clone Number | S-4102 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, IHC-P, ICC, ChIP |
| Reactivity | Hu, Mk |
| Positive Sample | HeLa, 293T, Ramos, COS-7 |
| Purification | Protein A |
| Concentration | 0.5 mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300 |
| Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied |
Dilution
| application | dilution | species |
| WB | 1:1000-1:2000 | Hu, Mk |
| IHC-P | 1:2000 | Hu |
| ICC | 1:500 | Hu |
| ChIP | 1:20-1:50 | Hu |
Background
SP1 (specificity protein 1) is a ubiquitous, C2H2-type zinc-finger transcription factor that binds GC-rich motifs (GGGCGG and related sequences) in the promoters and enhancers of thousands of human genes via its three C-terminal zinc fingers, recruiting TFIID, TFIIB, histone acetyl-transferases (p300/CBP, PCAF) and chromatin-remodeling complexes to activate or fine-tune basal and signal-dependent transcription of housekeeping, metabolic, cell-cycle, angiogenic and differentiation genes; it is tightly regulated through phosphorylation by multiple kinases (ERK, AKT, DNA-PK, CDK), acetylation, SUMOylation, prolyl-isomerization and redox status of its two critical cysteines, and its activity is modulated by oncogenic, nutrient and stress signaling, making SP1 a central hub in cancer, diabetes, cardiovascular and neurodegenerative diseases, with elevated expression often correlating with poor prognosis, while synthetic mithramycin, chromomycin and novel zinc-finger inhibitors are explored to target its DNA-binding or protein–protein interfaces therapeutically.
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Picture
Western Blot
WB result of SP1 Recombinant Rabbit mAb
Primary antibody: SP1 Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: COS-7 whole cell lysate 20 µg
Secondary antibody: Goat Anti- rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 81 kDa
Observed MW: 100 kDa
Immunohistochemistry
IHC shows positive staining in paraffin-embedded human breast cancer. Anti-SP1 antibody was used at 1/2000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human gastric cancer. Anti-SP1 antibody was used at 1/2000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
Immunocytochemistry
ICC shows positive staining in HeLa cells. Anti-SP1 antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 4% PFA and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).
ChIP
Chromatin immunoprecipitation (ChIP) was performed on HeLa cells cross - linked with 1% formaldehyde for 10 min, then chromatin was fragmented by sonication. Parallel reactions used SP1 Recombinant Rabbit mAb (S-4102) and Rabbit mAb IgG Isotype Control (SDT-R173) at 1:50 for immunoprecipitation.
Post -immunoprecipitation, both samples were washed, eluted, and cross - links reversed. Purified DNA was analyzed by qPCR.
qPCR showed the enrichment of DHFR, TNIP1 and SAT-α in SP1 Recombinant
Rabbit mAb (S-4102) -immunoprecipitated sample.
