RACK1 GNB2L1 Antibody

SKU: orb234289

Description

Anti-RACK1 GNB2L1 Antibody. Tested in Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.

Images & Validation

• Tested Applications: FC, ICC, IF, WB
• Reactivity: Human, Mouse, Rat
• Application Notes:
  Western blot, 0.1-0.5μg/ml, Human, Mouse, Rat Immunocytochemistry/Immunofluorescence, 2μg/ml, Human Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human. Add 0.2ml of distilled water will yield a concentration of 500ug/ml

Key Properties

• Antibody Type: Primary Antibody
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: Rabbit IgG
• Immunogen: E.coli-derived human RACK1 recombinant protein (Position: T2-R317). Human RACK1 shares 100% amino acid (aa) sequence identity with both mouse and rat RACK1.
• Molecular Weight: 32 kDa
• Purification: Immunogen affinity purified.

Storage & Handling

• 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.
• Form/Appearance: Lyophilized
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Disclaimer: For research use only

Alternative Names

Guanine nucleotide-binding protein subunit beta-2-like 1; Cell proliferation-inducing gene 21 protein; Guanine nucleotide-binding protein subunit beta-like protein 12.3; Human lung cancer oncogene 7 protein; HLC-7; Receptor for activated C kinase; Receptor of activated protein kinase C 1; RACK1; Guanine nucleotide-binding protein subunit beta-2-like 1, N-terminally processed; GNB2L1; HLC7, PIG21

Flow Cytometry analysis of A431 cells using anti-RACK1 antibody. Overlay histogram showing A431 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-RACK1 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.

IF analysis of RACK1 using anti-RACK1 antibody. RACK1 was detected in immunocytochemical section of A431 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2 µg/mL rabbit anti-RACK1 Antibody overnight at 4°C. DyLight®550 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Western blot analysis of RACK1 using anti-RACK1 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 30 ug of sample under reducing conditions. Lane 1: human MCF-7 whole cell lysates, Lane 2: human Jurkat whole cell lysates, Lane 3: human HepG2 whole cell lysates, Lane 4: human A549 whole cell lysates, Lane 5: human Raji whole cell lysates, Lane 6: rat liver tissue lysates, Lane 7: mouse RAW264.7 whole cell lysates, Lane 8: mouse HEPA1-6 whole cell lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA 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-RACK1 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 1:5000 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 RACK1 at approximately 36 kDa and for RACK1 mature form at approximately 32 kDa. The expected band size for RACK1 is at 35 kDa.

Quick Database Links

UniProt

P63244