PA1-21016
antibody from Invitrogen Antibodies
Targeting: MAPK1
ERK, ERK2, MAPK2, p41mapk, PRKM1, PRKM2
Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [1]
- Immunohistochemistry [1]
- Chromatin Immunoprecipitation [1]
- Other assay [1]
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Validation data
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- Product number
- PA1-21016 - Provider product page
- 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
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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).
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).
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.
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.