Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Western Blot, Simple Western

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # 1013D
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Product Specifications

Immunogen

Phosphopeptide containing the human MSK1(S376) site.
Accession # O75582

Specificity

Detects human MSK1 and MSK2 when phosphorylated at S376 and S360, respectively.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for Human Phospho-MSK1(S376)/MSK2(S360) Antibody

Detection of Human Phospho-MSK1 (S376)/MSK2 (S360) antibody by Western Blot.

Detection of Human Phospho-MSK1 (S376)/MSK2 (S360) by Western Blot.

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.

Detection of Human MSK1/MSK2 by Simple WesternTM.

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.
Detection of Human MSK1/MSK2 by Western Blot

Detection of Human MSK1/MSK2 by Western Blot

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 (https://pubmed.ncbi.nlm.nih.gov/34133426), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human MSK1/MSK2 by Western Blot

Detection of Human MSK1/MSK2 by Western Blot

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 (https://pubmed.ncbi.nlm.nih.gov/34133426), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human MSK1/MSK2 by Western Blot

Detection of Human MSK1/MSK2 by Western Blot

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 (https://pubmed.ncbi.nlm.nih.gov/34133426), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human Phospho-MSK1(S376)/MSK2(S360) Antibody

Application
Recommended Usage

Simple Western

5 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line treated PMA 

Western Blot

0.1 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line treated with PMA and Recombinant Human EGF (Catalog # 236-EG)

Formulation, Preparation, and Storage

Purification

Protein A or G purified from cell culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.


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Formulation

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.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & 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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: MSK1/MSK2

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. 

Long Name

Mitogen- and Stress-activated Protein Kinase 1 and 2

Alternate Names

1110069D02Rik, 3110005L17Rik, 6330404E13Rik

UniProt

Additional MSK1/MSK2 Products

Product Documents for Human Phospho-MSK1(S376)/MSK2(S360) Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human Phospho-MSK1(S376)/MSK2(S360) Antibody

For research use only

Citations for Human Phospho-MSK1(S376)/MSK2(S360) Antibody

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