Positive WB detected in: Hela whole cell lysate, MCF-7 whole cell lysate, 293 whole cell lysate, HepG2 whole cell lysate, Jurkat whole cell lysate, K562 whole cell lysate
All lanes: ATF4 antibody at 1.6μg/ml
Goat polyclonal to rabbit IgG at 1/50000 dilution
Predicted band size: 39 KDa
Observed band size: 50 KDa
IHC image of CSB-RA002272A0HU diluted at 1:160 and staining in paraffin-embedded human breast cancer performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a biotinylated secondary antibody and visualized using an HRP conjugated SP system.
Immunofluorescence staining of HepG2 cells with CSB-RA002272A0HU at 1:53, counter-stained with DAPI. The cells were fixed in 4% formaldehyde, permeabilized using 0.2% Triton X-100 and blocked in 10% normal Goat Serum. The cells were then incubated with the antibody overnight at 4°C. The secondary antibody was Alexa Fluor 488-congugated AffiniPure Goat Anti-Rabbit IgG (H+L).
Immunoprecipitating ATF4 in Hela whole cell lysate
Lane 1: Rabbit control IgG instead of CSB-RA002272A0HU in Hela whole cell lysate.
For western blotting, a HRP-conjugated Protein G antibody was used as the secondary antibody (1/2000)
Lane 2: CSB-RA002272A0HU (3μg) + Hela whole cell lysate (500μg)
Lane 3: Hela whole cell lysate (20μg)
Overlay histogram showing Hela cells stained with CSB-RA002272A0HU (red line) at 1:50. The cells were fixed with 70% Ethylalcohol (18h) and then permeabilized with 0.3% Triton X-100 for 2 min. The cells were then incubated in 1x PBS /10% normal goat serum to block non-specific protein-protein interactions followed by primary antibody for 1 h at 4°C. The secondary antibody used was FITC goat anti-rabbit IgG (H+L) at 1/200 dilution for 1 h at 4°C. Control antibody (green line) was used under the same conditions. Acquisition of >10,000 events was performed.
Transcriptional activator. Binds the cAMP response element (CRE) (consensus: 5'-GTGACGT[AC][AG]-3'), a sequence present in many viral and cellular promoters. Cooperates with FOXO1 in osteoblasts to regulate glucose homeostasis through suppression of beta-cell production and decrease in insulin production (By similarity). It binds to a Tax-responsive enhancer element in the long terminal repeat of HTLV-I. Regulates the induction of DDIT3/CHOP and asparagine synthetase (ASNS) in response to endoplasmic reticulum (ER) stress. In concert with DDIT3/CHOP, activates the transcription of TRIB3 and promotes ER stress-induced neuronal apoptosis by regulating the transcriptional induction of BBC3/PUMA. Activates transcription of SIRT4. Regulates the circadian expression of the core clock component PER2 and the serotonin transporter SLC6A4. Binds in a circadian time-dependent manner to the cAMP response elements (CRE) in the SLC6A4 and PER2 promoters and periodically activates the transcription of these genes. During ER stress response, activates the transcription of NLRP1, possibly in concert with other factors (PubMed:26086088).
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.
Cyclic AMP-dependent transcription factor ATF-4, cAMP-dependent transcription factor ATF-4, Activating transcription factor 4, Cyclic AMP-responsive element-binding protein 2, CREB-2, cAMP-responsive element-binding protein 2, DNA-binding protein TAXREB67, Tax-responsive enhancer element-binding protein 67, TaxREB67, ATF4, CREB2, TXREB
Epigenetics and Nuclear Signaling
Upon receipt, store at -20°C or -80°C. Avoid repeated freeze.