Product Name: ERK5-pY221
Product Number: AB-PK626
Size: 25 µg      Price:89.00
      $US
Target Full Name: Extracellular regulated protein-serine kinase 5 (Big MAP kinase 1 (BMK1))

Target Alias: BMK; BMK1; BMK1 kinase; ERK4; ERK-5; Extracellular-signal-regulated kinase 5; Kinase ERK5; MAPK7; PRKM7; ENSG00000166484

Product Type Specific: Protein kinase phosphosite-specific antibody

Antibody Code: PK626

Antibody Target Type: Phosphosite-specific

Antibody Phosphosite: Y221

Protein UniProt: Q13164

Protein SigNET: Q13164

Antibody Type: Polyclonal

Antibody Host Species: Rabbit

Antibody Immunogen Source: Human ERK5 (MAPK7) sequence peptide Cat. No.: PE-04AEI99

Antibody Immunogen Sequence: MTE(pY)VAT(bA)C

Antibody Immunogen Description: Corresponds to amino acid residues M218 to T224; In the protein kinase catalytic domain activation T loop region between subdomains VII and VIII.

Production Method: The immunizing peptide was produced by solid phase synthesis on a multipep peptide synthesizer and purified by reverse-phase hplc chromatography. Purity was assessed by analytical hplc and the amino acid sequence confirmed by mass spectrometry analysis. This peptide was coupled to KLH prior to immunization into rabbits. New Zealand White rabbits were subcutaneously injected with KLH-coupled immunizing peptide every 4 weeks for 4 months. The sera from these animals was applied onto an agarose column to which the immunogen peptide was thio-linked. Antibody was eluted from the column with 0.1 M glycine, pH 2.5. Subsequently, the antibody solution was neutralized to pH 7.0 with saturated Tris.This antibody was also subject to negative purification over phosphotyrosine-agarose.

Antibody Modification: Unconjugated. Contact KInexus if you are interest in having the antibody biotinylated or coupled with fluorescent dyes.

Antibody Concentration: 1 mg/ml

Storage Buffer: Phosphate buffered saline pH 7.4, 0.05% Thimerasol

Storage Conditions: For long term storage, keep frozen at -40°C or lower. Stock solution can be kept at +4°C for more than 3 months. Avoid repeated freeze-thaw cycles.

Product Use: Western blotting | Antibody microarray

Antibody Dilution Recommended: 2 µg/ml for immunoblotting

Antibody Potency: Medium immunoreactivity of a target-sized protein by Western blotting in A431 cells. Medium immunoreactivity with immunogen peptide on dot blots.

Antibody Species Reactivity: Human

Antibody Positive Control: The observed molecular mass of the processed target protein on SDS-PAGE gels is reported to be around 88-115 kDa.

Antibody Specificity: Medium-High

Antibody Cross Reactivity: No significant cross-reactive proteins detected in T98G cells, but manycross-reactive proteins detected in A431 cells, including phospho-ERK and phospho-p38 MAP kinases.

Related Product 1: ERK5-pY221 blocking peptide

Related Product 2: ERK5-2 pan-specific antibody (Cat. No.: AB-NK206-4)

Related Product 3: ERK5-3 pan-specific antibody (Cat. No.: AB-NK206-5)

Related Product 4: ERK5-NT (ERK5-4) pan-specific antibody (Cat. No.: AB-NK206-2)

Scientific Background: ERK5 (MAPK7) is a protein-serine/threonine kinase of the CMGC group and MAPK family. It is important in controlling a cell's proliferation, differentiation, and cell survival. Phosphorylation of T219 and Y221 by MEK5 (MAP2K5) increases the phosphotransferase activity of ERK5. MEK5 is activated in response to hyperosmolarity, hydrogen peroxide, and epidermal growth factor (EGF). ERK5 may be a tumour requiring protein (TRP). Although there has been no direct linkage of ERK5 to formation of carcinomas in humans in vivo, in vitro ERK5 over-expression has been shown to increase metastasis, and ERK5 inhibition has been shown to inhibit osteosarcoma cell motility. Upregulation of the MEK5 gene has been noted in breast, prostate, and liver cancer.
Figure 1. Dot blotting ERK5-pY221 antibody with recombinant purified proteins.


Figure 2. Epitope mapping of ERK5-pY221 antibody with similar phosphopeptides on dot blots.


Figure 3. Identification of phosphosites related to ERK5-pY221.