		Technical Data
E3452-59B	ERK5, phosphorylated (Thr218, Tyr220) (Extracellular Signal-regulated Kinase 5, ERK-5, Big MAP Kinase 1, BMK1, BMK-1, Mitogen-activated Protein Kinase 7, MAP Kinase 7, MAPK 7, MAPK7, PRKM7)

Description:
Extracellular signal-related protein kinase 5/big mitogen-activated protein kinase 1 (ERK5/BMK1) is an 88kD member of the MAP kinase superfamily that becomes fully activated when phosphorylated on threonine 218 and tyrosine 220 by MEK5. ERK5 is activated in response to certain growth factors and bioactive peptides, however the upstream activators are distinct from those that activate ERK1&2. Negatively preadsorbed using a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated ERK5. The final product is generated by affinity chromatography using an ERK5-derived peptide that is phosphorylated at threonine 218 and tyrosine 220.

Applications:
Suitable for use in Western Blot and Immunohistochemistry. Other applications not tested.

Recommended Dilutions:
Western Blot: 1:1000
Optimal dilutions to be determined by the researcher.

Positive Control Used:
HEK293 cells transiently co-transfected with plasmids expressing ERK5 kinase domain (ERK5kin) and constitutively activated MEK5 (MEK5D-D).

Storage and Stability:
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

Type	Isotype	Clone	Grade
Pab	IgG		Affinity Purified

Size		Storage	Shipping	Source	Host
10 blots		-20°C	Blue Ice	Human	Rabbit

Concentration:

Not Determined

Immunogen:

Synthetic phosphopeptide corresponding the region of human ERK5 that contains threonine 218 and tyrosine 220.

Purity:

Purified by immunoaffinity chromatography.

Form

Supplied as a liquid in PBS (without Mg2+ and Ca2+), pH 7.3, 1mg/ml BSA (IgG, protease free), 0.05% sodium azide, 50% glycerol. Also available without BSA & Azide. See E3452-59B1.

Specificity:

Recognizes human ERK5 when phosphorylated at Thr218 and Tyr220. Species sequence homology: mouse 100%. Due to the low abundance and low levels of activation of endogenous ERK5, overexpression or immunoprecipitation of ERK5 protein may be required to detect activation. Some cross-reactivity is observed with endogenous ERK1&2 (44 and 42kD, respectively) due to the high levels of expression and activation of this protein typically observed with most cell types.

Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.

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