ERK5 Recombinant Rabbit Monoclonal Antibody [SD2084] - BSA and Azide free
Catalog# HA750299
ERK5 Recombinant Rabbit Monoclonal Antibody [SD2084] - BSA and Azide free
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WB
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IF-Cell
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IF-Tissue
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IP
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FC
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Human
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Mouse
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Rat
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ET1612-7
含抗保成分
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unconjugated
Overview
Product Name
ERK5 Recombinant Rabbit Monoclonal Antibody [SD2084] - BSA and Azide free
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within Human ERK5 aa 767-816 / 816.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IF-Cell, IF-Tissue, IP, FC
Molecular Weight
Predicted band size: 88 kDa
Positive Control
HeLa cell lysate, NIH/3T3 cell lysate, Neuro-2a cell lysate, mouse brain tissue lysate, mouse testis tissue lysate, PC-12 cell lysate, NIH/3T3, PC-12, HeLa.
Conjugation
unconjugated
Clone Number
SD2084
Product Features
Form
Liquid
Concentration
Storage Instructions
Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage Buffer
1*PBS (pH7.4).
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:500-1:2,000
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IF-Cell
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1:100
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IF-Tissue
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1:100-1:500
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FC
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1:1,000
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IP
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Use at an assay dependent concentration.
Target
Function
The activation of signal transduction pathways by growth factors, hormones and neurotransmitters is mediated through two closely related MAP kinases, p44 and p42, designated extracellular-signal related kinase 1 (ERK 1) and ERK 2, respectively. ERK proteins are regulated by dual phosphorylation at specific tyrosine and threonine sites mapping within a characteristic Thr-Glu-Tyr motif. Phosphorylation at both the Thr and Tyr residues is required for full enzymatic activation. In response to activation, MAP kinases phosphorylate downstream components on serine and threonine. Upstream MAP kinase regulators include MAP kinase kinase (MEK), MEK kinase and Raf-1. The ERK family has three additional members: ERK 3, ERK 5 and ERK 6.
Background References
1. Rovida, E. et al. 2015. The mitogen-activated protein kinase ERK5 regulates the development and growth of hepatocellular carcinoma. Gut. 64: 1454-65.
2. Yoshihara, D. et al. 2011. PPAR-gamma agonist ameliorates kidney and liver disease in an orthologous rat model of human autosomal recessive polycystic kidney disease. Am. J. Physiol. Renal Physiol.. 300: F465-F474.
Sequence Similarity
Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. MAP kinase subfamily.
Tissue Specificity
Expressed in many adult tissues. Abundant in heart, placenta, lung, kidney and skeletal muscle. Not detectable in liver.
Post-translational Modification
Dually phosphorylated on Thr-219 and Tyr-221, which activates the enzyme (By similarity). Autophosphorylated in vitro on threonine and tyrosine residues when the C-terminal part of the kinase, which could have a regulatory role, is absent.
Subcellular Location
Cytoplasm, Nucleus.
Synonyms
Big MAP kinase 1 antibody
BMK 1 antibody
BMK 1 kinase antibody
BMK-1 antibody
BMK1 antibody
BMK1 Kinase antibody
EC 2.7.11.24 antibody
ERK 4 antibody
ERK 5 antibody
ERK-5 antibody
ExpandImages
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Western blot analysis of ERK5 on different lysates with Rabbit anti-ERK5 antibody (HA750299) at 1/1,000 dilution.
Lane 1: HeLa cell lysate (20 µg/Lane)
Lane 2: NIH/3T3 cell lysate (20 µg/Lane)
Lane 3: Neuro-2a cell lysate (20 µg/Lane)
Lane 4: Mouse brain tissue lysate (40 µg/Lane)
Lane 5: Mouse testis tissue lysate (40 µg/Lane)
Lane 6: PC-12 cell lysate (20 µg/Lane)
Predicted band size: 88 kDa
Observed band size: 115 kDa
Exposure time: 3 minutes; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750299) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
☑ Knockdown (KD)
Western blot analysis of ERK5 on different lysates with Rabbit anti-ERK5 antibody (HA750299) at 1/2,000 dilution.
Lane 1: A549-si NT cell lysate
Lane 2: A549-si ERK5 cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 88 kDa
Observed band size: 115 kDa
Exposure time: 1 minute; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750299) at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Immunocytochemistry analysis of NIH/3T3 cells labeling ERK5 with Rabbit anti-ERK5 antibody (HA750299) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-ERK5 antibody (HA750299) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunocytochemistry analysis of PC-12 cells labeling ERK5 with Rabbit anti-ERK5 antibody (HA750299) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-ERK5 antibody (HA750299) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Flow cytometric analysis of HeLa cells labeling ERK5.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA750299, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). -
Flow cytometric analysis of NIH/3T3 cells labeling ERK5.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA750299, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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