Phospho-MSK1 (Ser212) Rabbit Polyclonal Antibody

• Catalog Number: orb6445
• Category: Antibodies
• Description: Phospho-MSK1 (Ser212) Rabbit Polyclonal Antibody
• Target: RPS6KA5
• Clonality: Polyclonal
• Species/Host: Rabbit
• Isotype: IgG
• Conjugation: Unconjugated
• Reactivity: Human, Rat
• Predicted Reactivity: Bovine, Canine, Equine, Gallus, Mouse, Porcine, Rabbit
• Form/Appearance: Liquid
• Concentration: 1mg/ml
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Immunogen: KLH conjugated synthesised phosphopeptide derived from human MSK1 around the phosphorylation site of Ser212 AY(p-S)FC
• UniProt ID: O75582
• RRID: AB_10926818
• MW: 90 kDa
• Tested applications: FC, ICC, IF, IHC-Fr, IHC-P, WB
• Dilution range: WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, ICC/IF=1:50, IF=1:100-500, Flow-Cyt=2ug/Test
• Antibody Type: Primary Antibody
• 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: MSK1(phospho S212); MSK1(phospho Ser212); 90 kDa r
• Note: For research use only

Blank control (black line): Hela. Primary Antibody (green line): Rabbit Anti-Phospho-MSK1 (Ser212) antibody (orb6445), dilution: 2 ug/Test, Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC, dilution: 0.5 ug/Test. Isotype control (orange line): Normal Rabbit IgG, Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at -20°C, The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed.

Hela cell, 4% Paraformaldehyde-fixed, Triton X-100 at room temperature for 20 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Antibody incubation with (Phospho-MSK1 (Ser212)) polyclonal Antibody, Unconjugated (orb6445) 1:50, 90 minutes at 37°C, followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.

Sample: Lane 1: Human 293T cell lysates, Lane 2: Human HeLa cell lysates, Lane 3: Human MCF-7 cell lysates, Lane 4: Human HUVEC cell lysates, Lane 5: Human A549 cell lysates, Lane 6: Human MDA-MB-231 cell lysates, Primary: Anti-Phospho-MSK1 (Ser212) (orb6445) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 90 kDa, Observed band size: 85 kDa.

Tissue/Cell: Rat brain tissue, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min, Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Incubation: Anti-phospho-MSK1 Polyclonal Antibody, Unconjugated (orb6445) 1:500, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.