PA5-17710

antibody from Invitrogen Antibodies

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

Antibody data

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Validation data

MAPK1 protein structure - PA5-17710 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on whole cell extracts (30 µg 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) and MCF7 (Lane 10). The blot was probed with Rabbit Anti-ERK2 Polyclonal Antibody (Product # PA5-17710, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/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 Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot of MAPK1 was performed by loading 30 µg of WT (lane 1) and MAPK1 CRISPR KO (lane 2) HeLa cell lysates in RIPA buffer onto a 4-15% gradient polyacrylamide gel. Proteins on the blots were visualized with Ponceau staining (below immunoblot). Proteins were transferred to nitrocellulose membrane and blocked in 5% milk for 1 hr. MAPK1 was detected at approximately 41 kDa using a MAPK1 polyclonal antibody (Product # PA5-17710) at a dilution of 1:1000 in 5% BSA in TBS with 0.1% Tween 20 (TBST) overnight at 4 deg. The peroxidase-conjugated secondary antibody (Product # 65-6120) was diluted to 0.2 µg/mL in TBST with 5% milk for 1 hr. Chemiluminescent detection was performed using Pierce ECL Western Blotting Substrate (Product # 32106). Data courtesy of YCharOS Inc., an open science company with the mission of characterizing commercially available antibodies using knockout validation.