Invitrogen

ERK5 Recombinant Rabbit Monoclonal Antibody (SD2084)

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This Antibody was verified by Knockdown to ensure that the antibody binds to the antigen stated. View Details

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Datasheet

Questions & Answers

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ERK5 Antibody in Immunocytochemistry (ICC) — Immunocytochemical analysis of ERK5 in MCF-7 cells using a ERK5 Monoclonal antibody (Product # MA5-32389) as seen in green. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

ERK5 Antibody in Western Blot (WB) — Western blot was performed using Anti-ERK5 Recombinant Rabbit Monoclonal Antibody (SD2084) (Product # MA5-32389) and a 115 kDa band corresponding to Mitogen-activated protein kinase 7 was observed across cell lines and tissue tested. Upon EGF and sorbitol treatment in HeLa cells, there was a shift in the band size which corresponds to phosphorylated form of ERK5, since it is an autophosphorylated protein. Whole cell extracts (30 µg lysate) of (Fig.a) HeLa (Lane 1), HeLa serum starved for overnight and treated with EGF (100 ng/mL for 15 min) (Lane 2), HeLa treated with Sorbitol (0.5M for 30 min) (Lane 3) were electrophoresed using NuPAGE™ 3-8% Tris-Acetate Protein Gel (Product # EA0378BOX); (Fig.b) PC-3 (Lane 1), DU 145 (Lane 2), NIH:OVCAR-3 (Lane 3), SK-O-V3 (Lane 4), NIH/3T3 (Lane 5) and Mouse Testis (Lane 6) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:6000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

ERK5 Antibody in Flow Cytometry (Flow) — Flow Cytometric analysis of ERK5 in A549 cells using a ERK5 Monoclonal Antibody (Product # MA5-32389) at a dilution of 1:50, as seen in red compared with an unlabelled control (cells without incubation with primary antibody; black). Alexa Fluor 488-conjugated goat anti rabbit IgG was used as the secondary antibody.

ERK5 Antibody in Knockdown — Antibody specificity was demonstrated by siRNA mediated knockdown of target protein. DU 145 cells were transfected with Mitogen-activated protein kinase 7 siRNA and decrease in signal intensity was observed in Western Blot application using Anti-ERK5 Recombinant Rabbit Monoclonal Antibody (SD2084) (Product # MA5-32389). {KD}

ERK5 Antibody in Cell treatment — Altered expression of proteins upon cell treatment demonstrates antibody specificity. Western blot using ERK5 Recombinant Rabbit Monoclonal Antibody (SD2084) (Product # MA5-32389), shows a shift in the band size upon EGF and sorbitol treatment in HeLa cells, which corresponds to phosphorylated form of ERK5, since it is an autophosphorylated protein. {TM}

Product Details

Applications

Tested Dilution

Publications

Flow Cytometry (Flow)

1:50-1:100

Immunocytochemistry (ICC)

1:100-1:500

Immunofluorescence (IF)

1:100-1:500

Western Blot (WB)

1:1,000-1:2,000

Product Specifications

Species

Human, Mouse, Rat

Expression System

Rabbit / IgG

Class

Recombinant Monoclonal

Type

Antibody

Clone

SD2084

Immunogen

Synthetic peptide within C-terminal human ERK5

Conjugate

Unconjugated

Form

Liquid

Concentration

1 mg/mL

Purification

Protein A

Storage buffer

TBS, pH 7.4, with 40% Glycerol, 0.05% BSA

Contains

0.05% sodium azide

Storage conditions

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

RRID

AB_2809669

Product Specific Information

Recombinant rabbit monoclonal antibodies are produced using in vitro expression systems. The expression systems are developed by cloning in the specific antibody DNA sequences from immunoreactive rabbits. Then, individual clones are screened to select the best candidates for production. The advantages of using recombinant rabbit monoclonal antibodies include: better specificity and sensitivity, lot-to-lot consistency, animal origin-free formulations, and broader immunoreactivity to diverse targets due to larger rabbit immune repertoire.

Target Information

ERK5/MAPK7 is a member of the MAP kinase family. MAP kinases act as an integration point for multiple biochemical signals, and are involved in a wide variety of cellular processes such as proliferation, differentiation, transcription regulation and development. This kinase is specifically activated by mitogen-activated protein kinase kinase 5. It is involved in the downstream signaling processes of various receptor molecules including receptor type kinases, and G protein-coupled receptors. In response to extracelluar signals, this kinase translocates to cell nucleus, where it regulates gene expression by phosphorylating, and activating different transcription factors. Four alternatively spliced transcript variants of this gene encoding two distinct isoforms have been reported. ERK5 is activated by tyrosine and threonine phosphorylation It is expressed in many adult tissues, abundantly in heart, placenta, lung, kidney and skeletal muscle, but is not detectable in liver.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

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Bioinformatics

Protein Aliases: Big MAP kinase 1; BMK-1; BMK1 kinase; BNhp-1; BNhp1 kinase; ERK-5; extracellular signal regulated kinase 5; Extracellular signal-regulated kinase 5; extracellular-signal-regulated kinase 5; MAP kinase 7; MAPK 7; mitogen activated protein kinase 7; Mitogen-activated protein kinase 7

Gene Aliases: BMK-1; BMK1; ERK-5; ERK4; ERK5; Erk5-T; MAPK7; PRKM7

UniProt ID: (Human) Q13164, (Mouse) Q9WVS8

Entrez Gene ID: (Human) 5598, (Mouse) 23939, (Rat) 114509

Molecular Function: kinase non-receptor serine/threonine protein kinase protein kinase transferase