ERK1/2 Recombinant antibody

ERK1/2 Uni-rAb^TM Recombinant Antibody for WB, IHC, IF/ICC, FC (Intra), ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat

Applications

WB, IHC, IF/ICC, FC (Intra), ELISA

Conjugate

Unconjugated

CloneNo.

4P8

Cat no : 83533-1-RR

Synonyms

ERK1, ERK, ERK2, ERK-1, EC:2.7.11.24

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Validation Data Gallery

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  WB analysis using 83533-1-RR

  Various lysates were subjected to SDS PAGE followed by western blot with 83533-1-RR (ERK1/2 antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

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  IHC staining of human cervical cancer using 83533-1-RR

  Immunohistochemical analysis of paraffin-embedded human cervical cancer tissue slide using 83533-1-RR (ERK1/2 antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human cervical cancer using 83533-1-RR

  Immunohistochemical analysis of paraffin-embedded human cervical cancer tissue slide using 83533-1-RR (ERK1/2 antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human intrahepatic cholangiocarcinoma using 83533-1-RR

  Immunohistochemical analysis of paraffin-embedded human intrahepatic cholangiocarcinoma tissue slide using 83533-1-RR (ERK1/2 antibody) at dilution of 1:200 (under 20x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human intrahepatic cholangiocarcinoma using 83533-1-RR

  Immunohistochemical analysis of paraffin-embedded human intrahepatic cholangiocarcinoma tissue slide using 83533-1-RR (ERK1/2 antibody) at dilution of 1:200 (under 20x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of HeLa using 83533-1-RR

  Immunofluorescent analysis of (-20°C Ethanol) fixed HeLa cells using ERK1/2 antibody (83533-1-RR, Clone: 4P8 ) at dilution of 1:300 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L) (SA00013-1).

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  FC experiment of HepG2 using 83533-1-RR

  1x10^6 HepG2 cells were intracellularly stained with 0.25 ug ERK1/2 Recombinant antibody (83533-1-RR, Clone:4P8) and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2)(red), or 0.25 ug Control Antibody (blue). Cells were fixed and permeabilized with Transcription Factor Staining Buffer Kit (PF00011).

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  Affinity and Kinetic Characterization of 83533-1-RR

  Biolayer interferometry (BLl) kinetic assays of 83533-1-RR against Human ERK1/2 were performed. The affinity constant is 17.8 nM.

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Tested Applications

• Positive WB detected in: MCF-7 cells, Jurkat cells, HEK-293 cells, HepG2 cells, C6 cells, NIH/3T3 cells, rat brain tissue
• Positive IHC detected in: human cervical cancer tissue, human intrahepatic cholangiocarcinoma tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: HeLa cells
• Positive FC (Intra) detected in: HepG2 cells

Recommended dilution

• Western Blot (WB): WB : 1:500-1:2000
• Immunohistochemistry (IHC): IHC : 1:50-1:500
• Immunofluorescence (IF)/ICC: IF/ICC : 1:150-1:600
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.25 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Product Information

The immunogen of 83533-1-RR is ERK1/2 Fusion Protein expressed in E. coli.

• Tested Reactivity: human, mouse, rat
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen: ERK1/2 fusion protein Ag20350
• Full Name: mitogen-activated protein kinase 3
• Calculated Molecular Weight: 43 kDa
• Observed Molecular Weight: 38-44 kDa
• GenBank Accession Number: BC013992
• Gene Symbol: ERK1
• Gene ID (NCBI): 5595
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purfication
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

ERK1 and ERK2 belongs to the protein kinase superfamily. It is involved in both the initiation and regulation of meiosis, mitosis, and postmitotic functions in differentiated cells by phosphorylating a number of transcription factors such as ELK-1. ERK1/2 catalized the reaction: ATP + a protein = ADP + a phosphoprotein. It is activated by tyrosine phosphorylation in response to INS and NGF. This antibody can recognize both ERK1 and ERK2 with the molecular mass of 38-44 kDa.