p90 RSK1 (phospho-S732) antibody

• Catalog Number: orb345670
• Category: Antibodies
• Description: p90 RSK1 (phospho-S732) antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, WB
• Reactivity: Human
• Isotype: IgG
• Immunogen: This affinity purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to a region near the C-terminal end of human RSK1 protein.
• Concentration: 1.51 mg/mL
• Dilution range: ELISA: 1:5000-1:50,000, WB: 1:1000-1:3000
• Form/Appearance: Liquid (sterile filtered)
• Purity: This product was affinity purified from monospecific antiserum by immunoaffinity chromatography using phosphorylated peptide coupled to agarose beads followed by solid phase adsorption against non phosphorylated peptide. This antibody is specific for human p90 RSK1 protein phosphorylated at S732. A BLAST analysis was used to suggest cross reactivity with p90 RSK1 from human, rat, mouse, horse, bovine, and dog based on 100% homology with the immunizing sequence. Partially cross reactivity may occur against opossum and chicken based on > 90% homology with the immunizing sequence. Cross reactivity with p90 RSK1 pS732 from other sources has not been determined.
• Conjugation: Unconjugated
• UniProt ID: Q15418
• NCBI: NP_001006666.1
• Storage: Store vial at -20° C or below prior to opening. This vial contains a relatively low volume of reagent (25 µL). To minimize loss of volume dilute 1:10 by adding 225 µL of the buffer stated above directly to the vial. Recap, mix thoroughly and briefly centrifuge to collect the volume at the bottom of the vial. Use this intermediate dilution when calculating final dilutions as recommended below. Store the vial at -20°C or below after dilution. Avoid cycles of freezing and thawing.
• Buffer/Preservatives: 0.01% (w/v) Sodium Azide
• Alternative names: rabbit anti-p90 RSK1 pS732 antibody, rabbit anti-R
• Note: For research use only
• Application notes: This affinity-purified antibody has been tested for use in ELISA and western blotting. Specific conditions for reactivity should be optimized by the end user. By western blot a band approximately 90 kDa in size corresponding to p90 RSK1 pS732 protein is expected in the appropriate cell lysate or extract.
• Expiration Date: 12 months from date of receipt.

Western Blot of Rabbit anti-p90 RSK1 antibody. Lane 1: unstimulated HEK293T cell lysates. Lane 2: EGF stimulated HEK293T cell lysates. Load: 35 µg per lane. Primary antibody: p90 RSK1 antibody at 1:1000 for overnight at 4°C. Secondary antibody: peroxidase conjugated secondary antibody and ECL. Block: 5% BLOTTO overnight at 4°C. Predicted/Observed size: 90 kDa, ~90 kDa for p90 RSK1. Other band(s): none.

Western blot using Biorbyt's affinity purified anti-p90 RSK1 pS732 antibody shows detection of a band ~90 kDa in size corresponding to phosphorylated p90 RSK1 (arrowhead) in EGF stimulated (lane 2) HEK293T cell lysates prepared from cells grown in the absence of serum for 12 h. No staining is observed in similarly prepared lysates derived from unstimulated (control) cells (lane 1). After transfer, the membrane was blocked overnight followed by reaction with the primary antibody at a 1:1000 dilution. Detection occurred using a peroxidase conjugated secondary antibody and ECL.

Western blot using Biorbyt's affinity purified anti-p90 RSK1 pS732 antibody. Lane 1-2: HEK293T (U) untreated or (T) treated with EGF. Lane 3-4: HEK293T (U) untreated or (T) treated with EGF. Lane 5-6: HEK293T (U) untreated or (T) treated with EGF. Load: 15 µg per lane. Actin used as a loading control. Blocking: 5% milk. Primary Antibody: Anti-RSK1-pS732 1:1000, 1:2000, or 1:3000 O/N. Secondary Antibody: Goat Anti-Rabbit IgG 1:5000 for 2 hours. Predicted Size: ~90 kDa in size corresponding to phosphorylated p90 RSK1 in EGF stimulated.