Human/Mouse/Rat ERK2 Antibody

Name: Human/Mouse/Rat ERK2 Antibody
Brand: R&D Systems
SKU: MAB1230
Price: 459 USD
Availability: InStock

R&D Systems | Catalog # MAB1230

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Product Specifications
Scientific Data
Applications
Preparation & Storage
Calculators
Background
Product Documents
Specific Notices
Research Areas

Key Product Details

Validated by

Knockout/Knockdown, Biological Validation

Species Reactivity

Human, Mouse, Rat

Applications

Knockout Validated, Immunohistochemistry, Western Blot, Immunocytochemistry, Simple Western

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG_2A Clone # 191801

View all ERK2 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human ERK2
Met1-Ser360
Accession # Q1HBJ4

Specificity

Detects human, rat and mouse ERK2. This antibody does not recognize endogenous ERK1.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG_2A

Scientific Data Images for Human/Mouse/Rat ERK2 Antibody

Detection of Human/Mouse/Rat ERK2 by Western Blot.

Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line, MCF-7 human breast cancer cell line, U937 human histiocytic lymphoma cell line, PC-12 rat adrenal pheochromocytoma cell line, and NIH-3T3 mouse embryonic fibroblast cell line. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human/Mouse/Rat ERK2 Monoclonal Antibody (Catalog # MAB1230) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF007). A specific band was detected for ERK2 at approximately 44 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

ERK2 in 3T3‑L1 Mouse Cell Line.

ERK2 was detected in immersion fixed 3T3-L1 mouse embryonic fibroblast adipose-like cell line using Mouse Anti-Human/Mouse/Rat ERK2 Monoclonal Antibody (Catalog # MAB1230) at 3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm and nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of Human ERK2 by Simple Western^TM.

Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line and MCF‑7 human breast cancer cell line, loaded at 0.5 mg/mL. A specific band was detected for ERK2 at approximately 43 kDa (as indicated) using 10 µg/mL of Mouse Anti-Human/Mouse/Rat ERK2 Monoclonal Antibody (Catalog # MAB1230). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Western Blot Shows Human ERK2 Specificity by Using Knockout Cell Line.

Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and ERK2 knockout HeLa cell line (KO). PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human/Mouse/Rat ERK2 Monoclonal Antibody (Catalog # MAB1230) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for ERK2 at approximately 42 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (Catalog # MAB5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Mouse ERK2 by Western Blot

Enhanced Dectin-1 Signaling in Fcer1g−/− DCs. Six-day-cultured BMDCs derived from WT and Fcer1g−/− mice were collected and starved in serum-free RPMI. After 3 h, cells were treated with dZym and lysed at indicated time points. The proteins were separated by SDS-PAGE, transferred to PVDF membranes, and then analyzed by Western blot. The detection of phosphorylated and total proteins of Src, Syk, AKT, PLC gamma, ERK, p38MAPK, and c-Raf (A), and total proteins of I kappa B alpha and GAPDH (B), were shown. Quantification was determined by densitometry using ImageJ software and the number represented the fold of each (phosphoprotein/total protein) value normalized to the (phosphoprotein/total protein) value of untreated WT control (WT 0 min). All data shown were representative from three to five independent experiments. Image collected and cropped by CiteAb from the following publication (https://journal.frontiersin.org/article/10.3389/fimmu.2017.01424/full), licensed under a CC-BY license. Not internally tested by R&D Systems.

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Applications for Human/Mouse/Rat ERK2 Antibody

Application

Recommended Usage

Immunocytochemistry

3-25 µg/mL
Sample: Immersion fixed 3T3-L1 mouse embryonic fibroblast adipose-like cell line

Immunohistochemistry

8-25 µg/mL
Sample: Perfusion fixed frozen sections of rat brain (cortex)

Knockout Validated

ERK2 is specifically detected in HeLa human cervical epithelial carcinoma parental cell line but is not detectable in ERK2 knockout HeLa cell line.

Simple Western

10 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line and MCF‑7 human breast cancer cell line

Western Blot

1 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line, MCF-7 human breast cancer cell line, U937 human histiocytic lymphoma cell line, PC-12 rat adrenal pheochromocytoma cell line, and NIH-3T3 mouse embryonic fibroblast cell line

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

Volume (to add to vial)

Mass (in vial)

Desired Reconstitution Concentration

Background: ERK2

ERK1 and ERK2 (also known as MAPK3 and MAPK1) are 44 and 42 kDa Ser/Thr kinases, respectively. They are part of the Ras-Raf-ERK signal transduction cascade often found downstream of growth factor receptor activation. ERK1 and ERK2 were initially isolated and cloned as kinases activated in response to insulin and NGF. They are expressed in most, if not all, mammalian tissues. Dual threonine and tyrosine phosphorylation activate both ERKs, at Thr202/Tyr204 for human ERK1 and Thr185/Tyr187 for human ERK2.

ERK5, also known as Big Mitogen-activated Protein Kinase 1 (BMK1) and MAPK7, is activated by several mechanisms, including receptor tyrosine kinases, G protein-coupled receptors, and osmotic stress. Like ERK1 and ERK2, ERK5 contains the conserved Thr-Glu-Tyr activation motif in its activation loop. Unlike these ERKs, however, ERK5 contains a unique C-terminal domain that regulates its activation and nuclear translocation.