MSK1/MSK2 [p Ser376, p Ser360] Antibody (1013D) [Unconjugated]

Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line untreated (-) or treated (+) with 200 nM PMA for 20 minutes and 100 ng/mL Recombinant Human EGF (Catalog # 236-EG) for 5 minutes. PVDF membrane was probed with 0.1 µg/mL of Rabbit Anti-Human Phospho-MSK1/MSK2 (MSK1 S376, MSK2 S360) Monoclonal Antibody (Catalog # MAB1094) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Phospho-MSK1 (S376)/MSK2 (S360) at approximately 95 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line untreated (-) or treated (+) with 200 nm PMA for 20 minutes, loaded at 0.2 mg/mL. A specific band was detected for MSK1/MSK2 at approximately 98 kDa (as indicated) using 5 µg/mL of Rabbit Anti-Human Phospho-MSK1/MSK2 (MSK1 S376, MSK2 S360) Monoclonal Antibody (Catalog # MAB1094) . This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Time course of phosphorylation changes after clofarabine treatment of STA-ET-7.2 cells.(A) STA-ET-7.2 cells treated with 0.5μM clofarabine and total cell lysates collected at different time points. Western blot results showed increased phosphorylation starting as early as 3 minutes; (B) phosphorylation changes were sustained through 25 hours with a reduction around one hour. Actin was used as loading control. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/34133426), licensed under a CC-BY license. Not internally tested by R&D Systems.

Phosphorylation changes after treatment for different ES and OS cell lines.(A) Cytarabine’s effect on STA-ET.7.2 cells compared to the clofarabine treatment. Cells were treated with clofarabine, cytarabine or DMSO. Immunoblot analysis showed cytarabine did not alter phosphorylation of ERK1/2, MSK1/2 and CREB; (B) Immunoblot results of phosphorylation changes after clofarabine treatment of different ES cell lines; clofarabine caused increased phosphorylation of ERK1/2, MSK1/2 and CREB in all five ES cell lines; (C) Clofarabine treated OS cell lines did not show significant increase in phosphorylation of ERK1/2, MSK1/2 and CREB. Actin was used as the loading control for all experiments. (CLF: Clofarabine, CYT: Cytarabine). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/34133426), licensed under a CC-BY license. Not internally tested by R&D Systems.

Clofarabine and CD99 antibodies cause specific protein phosphorylation in ES cell lines.(A) STA-ET-7.2 cells treated with clofarabine and western blot analyses were done by phosphospecific antibodies. Phosphorylation of ERK1/2, MSK1/2 and CREB levels were increased with clofarabine treatment; (B) TC-71 cells treated with two different C99 antibodies (15μg/ml) and phosphorylation changes were analyzed by immunoblotting. Similar to clofarabine treatment, phosphorylation of ERK1/2, MSK1/2 and CREB were increased; (C, D) STA-ET-7.2 cells treated with clofarabine, CD 99 antibody and decreased phosphorylation changes for STAT3 and AMPK alpha 1 were validated by western blotting; (E) A4573 cells treated with clofarabine show increased level of pMSK1/2 and main phosphorylated protein is pMSK2. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/34133426), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: WB, Simple Western
• Clone: 1013D
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated

MSK1/MSK2 [p Ser376, p Ser360] Antibody (1013D) [Unconjugated] Summary

• Additional Information: Recombinant Monoclonal Antibody.
• Immunogen: Phosphopeptide containing the human MSK1(S376) site.
  Accession # O75582
• Modification: p Ser376, p Ser360
• Specificity: Detects human MSK1 and MSK2 when phosphorylated at S376 and S360, respectively.
• Source: N/A
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Purity Statement: Protein A or G purified from cell culture supernatant

Applications/Dilutions

• Dilutions:
  • Simple Western 5 ug/mL
  • Western Blot 0.1 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: Sodium Azide
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for MSK1/MSK2 [p Ser376, p Ser360] Antibody (1013D) [Unconjugated]

• 1110069D02Rik
• 3110005L17Rik
• 6330404E13Rik
• MSK1/MSK2

Background

Mitogen- and Stress-activated protein Kinases 1 and 2 (MSK1/2) have been shown to play key roles in the transcriptional regulation of immediate early genes such as c-fos. MSK1, also known as Ribosomal Protein S6 Kinase 5 (RPS6KA5), and MSK2, also known as RSKB and RPS6KA4, belong to the AGC family of kinases. Both proteins have two kinase domains connected by a regulatory linker region, and are activated by the mitogen-activated protein kinases ERK1, ERK2, and p38. Nuclear MSK phosphorylates and activates a number of transcription factors, including ATF1 and CREB. The phosphorylation of MSK1 at Ser376 or the equivalent Ser360 in MSK2 is required for kinase activity. These sites are located in the AGC kinase domain and are autophosphorylated. Their phosphorylation is essential for the catalytic activity of the N-terminal kinase domain. The sequence surrounding MSK1(S376) and MSK2(S360) is 100% identical. 

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for MSK1/MSK2 Antibody (MAB1094)(1)

Publication using MAB1094

• L Gresseau, ME Roy, S Duhamel, B Annabi A Signaling Crosstalk Links SNAIL to the 37/67 kDa Laminin-1 Receptor Ribosomal Protein SA and Regulates the Acquisition of a Cancer Stem Cell Molecular Signature in U87 Glioblastoma Neurospheres Cancers, 2022-11-30;14(23):. 2022-11-30 [PMID: 36497426] (Western Blot, Human)

Bioinformatics

• Uniprot:
  • O75582()