LF-MA0134

antibody from Invitrogen Antibodies

Targeting: MAPK3 ERK1, p44erk1, p44mapk, PRKM3

Western blot (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

LF-MA0134

Provider

Invitrogen Antibodies

Product name

Anti-ERK1/ERK2 Monoclonal Antibody (9B3)

Antibody type

Monoclonal

Antigen

Synthetic peptide

Description

A suggested positive control for this product is HepG2 cells.

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

9B3

Vial size

100 µL

Concentration

1 mg/mL

Storage

-20° C, Avoid Freeze/Thaw Cycles

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on whole cell extracts (30 µg lysate) of HEK-293 (Lane 1), A549 (Lane 2), HeLa (Lane 3), MCF7 (Lane 4), U-937 (Lane 5), HEL 92.1.7 (Lane 6), K-562 (Lane 7), A-431 (Lane 8), Jurkat (Lane 9), HT-29 (Lane 10) and HCT 116 (Lane 11). The blot was probed with ERK1/ERK2 Mouse monoclonal Antibody (Product # LF-MA0134, 2 µg/mL) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/mL, 1:4000 dilution). A 42 kDa band corresponding to ERK1 was observed across the cell lines tested and 44 kDa band corresponding to ERK2 was observed in HEK-293 and MCF7 cell lines. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX), 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 Pierce™ ECL Western Blotting Substrate (Product # 32106).

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry analysis of p44/42 MAP Kinase was done on HCT 116 cells. Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Triton™ X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with p44/42 MAP Kinase Mouse Monoclonal Antibody (Product # LF-MA0134, red histogram) or with mouse isotype control (pink histogram) at 3-5 µg/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor® 488 Rabbit Anti-Mouse Secondary Antibody (Product # A-11059) at a dilution of 1:400 for 30 minutes at room temperature. The representative 10,000 cells were acquired and analyzed for each sample using an Attune® Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.