PA1-21016

antibody from Invitrogen Antibodies

Targeting: MAPK1 ERK, ERK2, MAPK2, p41mapk, PRKM1, PRKM2

Western blot (Supportive data in Antibodypedia)

Immunoprecipitation (Recommended by provider)

Immunohistochemistry (Supportive data in Antibodypedia)

Chromatin Immunoprecipitation (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA1-21016

Provider

Invitrogen Antibodies

Product name

Anti-ERK2 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

PA1-21016 detects ERK2 in Human, Mouse and Rat samples. PA1-21016 has been successfully used in Immunohistochemistry (Paraffin), ChIP, Immunoprecipitation and Western Blot procedures. PA1-21016 immunogen corresponds to Synthetic peptide corresponding to the C-terminal region of the mouse protein.

Reactivity

Human, Mouse, Rat

Host

Rabbit

Isotype

IgG

Vial size

500 µL

Concentration

0.2 mg/ml

Storage

4° C, do not freeze

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on whole cell extracts (30 ug lysate) of NIH/3T3 (Lane 1), C2C12 (Lane 2), HeLa (Lane 3), H1975 (Lane 4), A-431 (Lane 5), A549 (Lane 6), HCT 116 (Lane 7), HT-29 (Lane 8), MDA-MB-231 (Lane 9), MCF7 (Lane 10) and Raji (Lane11). The blot was probed with Rabbit Anti-ERK2 Polyclonal Antibody (Product # PA1-21016, 1 ug/ml) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 ug/ml, 1:4000 dilution). A 42 kDa band corresponding to ERK2 was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10 % Bis-Tris gel (Product # NP0302BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical analysis of p42 MAP Kinase/ERK2/MAPK1 in human breast carcinoma using a p42 MAP Kinase/ERK2/MAPK1 polyclonal antibody (Product # PA1-21016).

Chromatin Immunoprecipitation

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Chromatin immunoprecipitation analysis of MAPK1 was performed using cross-linked chromatin from 1 x 106 HCT116 colon carcinoma cells treated with serum for 0, 15, 30, and 60 minutes. Immunoprecipitation was performed using a multiplex microplate Matrix ChIP assay (see reference for Matrix ChIP protocol: http://www.ncbi.nlm.nih.gov/pubmed/22098709) with 1.0ul/100ul well volume of a MAPK1 polyclonal antibody (Product # PA1-21016). Chromatin aliquots from ~1 x 105 cells were used per ChIP pull-down. Quantitative PCR data were done in quadruplicate using 1ul of eluted DNA in 2ul SYBR real-time PCR reactions containing primers to amplify -15kb upstream of the Egr1 gene or exon-1 of Egr1. PCR calibration curves were generated for each primer pair from a dilution series of sheared total genomic DNA. Quantitation of immunoprecipitated chromatin is presented as signal relative to the total amount of input chromatin. Results represent the mean +/- SEM for three experiments. A schematic representation of the Egr-1 locus is shown above the data where boxes represent exons (black boxes = translated regions, white boxes = untranslated regions); the zigzag line represents an intron; and the straight line represents upstream sequence. Regions amplified by Egr-1 primers are represented by black bars. Data courtesy of the Innovators Program.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

IP-MS enrichment of ERK2 (LFQ intensity): ERK2 was enriched 63-fold from HEPG2 lysate compared to background proteins, using the optimized IP-MS workflow with Pierce MS-Compatible Magnetic IP Kit protein A/G (Product # 90409) and ERK2 antibody (Product # PA1-21016). The STRING database (www.string-db.org) was used to identify the protein interactor list. See more information on IP-MS verification of antibody selectivity.