WB result of ATF3 Recombinant Rabbit mAb
Primary antibody: ATF3 Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: Daudi whole cell lysate 20 µg
Lane 2: HepG2 whole cell lysate 20 µg
Lane 3: 293T whole cell lysate 20 µg
Lane 4: HeLa whole cell lysate 20 µg
Negative control: Daudi whole cell lysate
Secondary antibody: Goat Anti- rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 21 kDa
Observed MW: 25 kDa
ATF3 Recombinant Rabbit mAb, PBS Only (SDT-4100)
ATF3 Recombinant Rabbit mAb, PBS Only (SDT-4100)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | ATF3 |
| Synonyms | Cyclic AMP-dependent transcription factor ATF-3; cAMP-dependent transcription factor ATF-3; Activating transcription factor 3 |
| Location | Nucleus |
| Accession | P18847 |
| Clone Number | SDT-4100 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, IHC-P, ICC, ChIP |
| Reactivity | Hu |
| Positive Sample | HepG2, 293T, HeLa, PMA & LPS treated THP-1 cells |
| Purification | Protein A |
| Concentration | 1 mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS |
| Stability & Storage | 12 months from date of receipt / reconstitution, 4 °C as supplied |
Dilution
| application | dilution | species |
| WB | 1:1000 | Hu |
| IHC-P | 1:500 | Hu |
| ICC | 1:500 | Hu |
| ChIP | 1:20-1:50 | Hu |
Background
ATF3 (Activating Transcription Factor 3) is a 181-amino-acid stress-inducible bZIP transcription factor that binds DNA as homo- or heterodimer, recognized by the consensus TGACGTCA, and acts as a dual-function transcriptional regulator—repressing or activating target genes involved in apoptosis, inflammation, cell-cycle checkpoints, metabolic reprogramming and oncogenesis—rapidily up-regulated in response to hypoxia, DNA damage, cytokines, ER stress or mechanical strain, frequently overexpressed in cancers where it promotes survival, metastasis and chemoresistance yet can also function as a tumor suppressor depending on cellular context, and is subject to tight control via MAPK, NF-κB, p53 and multiple feedback loops, making it a critical node in integrating stress signals and determining cell fate.
Picture
Picture
Western Blot
Immunohistochemistry
IHC shows positive staining in paraffin-embedded human placenta. Anti-ATF3 antibody was used at 1/500 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-ATF3 antibody was used at 1/500 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 Hodgkin’s lymphoma. Anti-ATF3 antibody was used at 1/500 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.
Negative control: IHC shows negative staining in paraffin-embedded human stomach. Anti-ATF3 antibody was used at 1/500 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 analysis of THP-1 cells treated with PMA (80nM,16h), then +LPS (1ug/ml, 8h) (top panel) and untreated THP-1 cells (below panel). Anti-ATF3 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 ATF3 Recombinant Rabbit mAb (SDT-4100) 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 ATF3 and SAT-α in ATF3 Recombinant Rabbit
mAb (SDT-4100) -immunoprecipitated sample.
