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: K562 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: 90 kDa
SP1 Recombinant Rabbit mAb (S-3073-71)
SP1 Recombinant Rabbit mAb (S-3073-71)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | SP1 |
| Synonyms | Transcription factor Sp1; TSFP1 |
| Immunogen | Synthetic Peptide |
| Location | Cytoplasm, Nucleus |
| Accession | P08047 |
| Clone Number | S-3073-71 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, IHC-P, ICC, IF, ChIP |
| Reactivity | Hu, Ms, Rt, Mk |
| Positive Sample | HeLa, K562, Ramos, NIH/3T3, 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 | 1:1000-1:5000 | Hu, Ms, Mk |
| IHC-P | 1:2000 | Hu, Ms, Rt |
| ICC | 1:500 | Hu, Ms |
| IF | 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: NIH/3T3 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: 90 kDa
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: 90 kDa
Immunohistochemistry
IHC shows positive staining in paraffin-embedded human colon. 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 colon 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 mouse kidney. 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 rat colon. 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).
ICC shows positive staining in NIH/3T3 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 100% ice-cold methanol and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).
Immunofluorescence
IF shows positive staining in paraffin-embedded human ovarian cancer. Anti-SP1 antibody was used at 1/500 dilution (magenta) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 647) (S0B4005) was used as secondary antibody at 1/500 dilution. Counterstained with DAPI (Blue). Heat mediated antigen retrieval with EDTA buffer pH9.0 was performed before commencing with IF staining protocol.
IF shows positive staining in paraffin-embedded human colon cancer. Anti-SP1 antibody was used at 1/500 dilution (magenta) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 647) (S0B4005) was used as secondary antibody at 1/500 dilution. Counterstained with DAPI (Blue). Heat mediated antigen retrieval with EDTA buffer pH9.0 was performed before commencing with IF staining protocol.
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-3073-71) 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-3073-71) -immunoprecipitated sample.
