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Catalog Number | orb402265 |
---|---|
Category | Antibodies |
Description | Anti-PERK/EIF2AK3 Antibody. Tested in ELISA, Flow Cytometry, WB applications. This antibody reacts with Human, Mouse, Rat. |
Species/Host | Rabbit |
Clonality | Polyclonal |
Tested applications | ELISA, FC, WB |
Reactivity | Human, Mouse, Rat |
Isotype | Rabbit IgG |
Immunogen | E. coli-derived human PERK recombinant protein (Position: R222-Q334). |
Antibody Type | Primary Antibody |
Concentration | Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml. |
Form/Appearance | Lyophilized |
Conjugation | Unconjugated |
MW | 140 kDa |
UniProt ID | Q9NZJ5 |
Storage | Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles. |
Alternative names | Eukaryotic translation initiation factor 2-alpha k Read more... |
Note | For research use only |
Application notes | Western blot, 0.1-0.5μg/ml Flow Cytometry (Fixed), 1-3μg/1x106 cells ELISA, 0.1-0.5μg/ml. Add 0.2ml of distilled water will yield a concentration of 500ug/ml |
Expiration Date | 12 months from date of receipt. |
Flow Cytometry analysis of HepG2 cells using anti-PERK antibody. Overlay histogram showing HepG2 cells (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-PERK Antibody (1 µg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10 µg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 µg/1x10^6) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Western blot analysis of PERK using anti-PERK antibody. Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50 ug of sample under reducing conditions. Lane 1: human Hela whole cell lysates, Lane 2: human COLO-320 whole cell lysates, Lane 3: human A549 whole cell lysates, Lane 4: human SK-OV-3 whole cell lysates, Lane 5: Human A431 whole cell lysates, Lane 6: rat brain tissue lysates, Lane 7: mouse brain tissue lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-PERK antigen affinity purified polyclonal antibody at 0.5 µg/mL overnight at 4°C, then washed with TBS-0.1% Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for PERK at approximately 140KD. The expected band size for PERK is at 125KD.
ELISA, FC, WB | |
Human, Mouse, Rat | |
Rabbit | |
Polyclonal | |
iFluor647 |