Antibody data
- Antibody Data
- Antigen structure
- References [3]
- Comments [0]
- Validations
- Western blot [5]
- Immunocytochemistry [1]
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Validation data
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- Product number
- 13-3500 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- MEK1 Monoclonal Antibody (3D9)
- Antibody type
- Monoclonal
- Antigen
- Purifed from natural sources
- Reactivity
- Human, Mouse, Rat, Canine, Xenopus
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 3D9
- Vial size
- 100 µg
- Concentration
- 0.5 mg/mL
- Storage
- -20°C
Submitted references PTPIP51—A New RelA-tionship with the NFκB Signaling Pathway.
Expression of Galpha 13 (Q226L) induces P19 stem cells to primitive endoderm via MEKK1, 2, or 4.
An analysis of Mek1 signaling in cell proliferation and transformation.
Brobeil A, Kämmerer F, Tag C, Steger K, Gattenlöhner S, Wimmer M
Biomolecules 2015 Apr 16;5(2):485-504
Biomolecules 2015 Apr 16;5(2):485-504
Expression of Galpha 13 (Q226L) induces P19 stem cells to primitive endoderm via MEKK1, 2, or 4.
Wang HY, Kanungo J, Malbon CC
The Journal of biological chemistry 2002 Feb 1;277(5):3530-6
The Journal of biological chemistry 2002 Feb 1;277(5):3530-6
An analysis of Mek1 signaling in cell proliferation and transformation.
Greulich H, Erikson RL
The Journal of biological chemistry 1998 May 22;273(21):13280-8
The Journal of biological chemistry 1998 May 22;273(21):13280-8
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of MEK1 was performed by loading 30 µg of cell lysate of mouse thymocytes untreated (lane 1) and treated with 2 µM AZD6244 for 18 hr at 37C (lane 2) in 1xSDS sample buffer and SeeBlue Plus2 Pre-stained Protein Standard (Product # LC5925) onto a NuPAGE Novex 4-12% Bis-Tris Protein Gels (Product #NP0324BOX). Proteins were transferred to nitrocellulose membrane of Trans-Blot Turbo Mini Nitrocellulose Transfer Packs using the Trans-Blot Turbo at 15-min 1.3A setting. Membrane was blocked in 1xDPBS (Product #14190136) containing 0.05% Tween 20 and 50% SeaBlock buffer (Product # 37527) for one hour at room temperature. MEK1 was detected at 43kDa molecular weights using a MEK1 antibody (Product # 13-3500) at a dilution of 1:1,000 in 1xDPBS (Product # 14190136) containing 0.05% Tween 20 and 50% SeaBlock buffer for one hour at room temperature on a rocking platform, followed by a Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody conjugated with Alexa Fluor Plus 680 (Product# A32729) at a dilution of 1:10,000 for one hour at room temperature. Fluorescent detection was performed using LI-COR Odyssey imager.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on whole cell extracts (30 µg lysate) of HeLa (Lane 1), H-1975 (Lane 2), HCT 116 (Lane 3), HT-29 (Lane 4), A-431 (Lane 5), A549 (Lane 6), MDA-MB-231 (Lane 7), U-87 MG (Lane 8), MCF7 (Lane 9), Raji (Lane 10) and NIH/3T3 (Lane 11). The blot was probed with Anti-MEK1 Mouse Monoclonal Antibody (Product # 13-3500, 1 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 45 kDa band corresponding to MEK1 was detected across all cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), 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).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of MEK1 was performed by loading 20 µg of the indicated whole cell lysates per well and 5 µL of PageRuler Plus Prestained Protein Ladder (Product # 26619) onto a 4-20% Tris-Glycine polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane using the G2 Blotter (Product # 62288), and blocked with 5% milk in TBST for 1 hour at room temperature. MEK1 was detected at ~43 kD using a MEK1 monoclonal antibody (Product # 13-3500) at a dilution of 1 µg/mL in 5% milk in TBST overnight at 4C on a rocking platform. After washing, blots were probed with a goat anti-mouse IgG-HRP secondary antibody (Product # 31430) at a dilution of 1:10,000 for at least 30 minutes at room temperature. Chemiluminescent detection was performed using SuperSignal West Pico substrate (Product # 34080).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockout of MEK1 was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR934388_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of MEK1 was performed by loading 30 µg of HeLa Cas9 (Lane 1), andHeLa MEK1 KO (Lane 2) whole cell extracts. The samples were electrophoresed using NuPAGE™ Novex™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with Anti-MEK1 Monoclonal Antibody (3D9) (Product # 13-3500, 1:1,000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:5,000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to MEK1.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of MEK1 was performed by loading 30 µg of cell lysate of mouse thymocytes untreated (lane 1) and treated with 2 µM AZD6244 for 18 hr at 37C (lane 2) in 1xSDS sample buffer and SeeBlue® Plus2 Pre-stained Protein Standard (Product # LC5925) onto a NuPAGE™ Novex™ 4-12% Bis-Tris Protein Gels (Product #NP0324BOX). Proteins were transferred to nitrocellulose membrane and blocked in 1xDPBS (Product #14190136) containing 0.05% Tween 20 and 50% SeaBlock buffer (Product # 37527) for one hour at room temperature. MEK1 was detected at 43kDa molecular weights using a MEK1 antibody (Product # 13-3500) at a dilution of 1:1,000 in 1xDPBS (Product # 14190136) containing 0.05% Tween 20 and 50% SeaBlock buffer for one hour at room temperature on a rocking platform, followed by a Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody conjugated with Alexa Fluor Plus 680 (Product # A32729) at a dilution of 1:10,000 for one hour at room temperature. Data courtesy of Ab Data Exchange Program.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of MEK1 was done on 70% confluent log phase MCF7 cells. The cells were fixed with 4% paraformaldehyde for 15 minutes; permeabilized with 0.25% Triton™ X-100 for 10 minutes followed by blocking with 5% BSA for 1 hour at room temperature. The cells were incubated with MEK1 Mouse Monoclonal Antibody (Product # 13-3500) at 2 µg - 4 µg in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Fluor® 488 Rabbit Anti-Mouse IgG Secondary Antibody (Product # A-11059) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 594 Phalloidin (Product # A12381). Panel d is a merged image showing cytoplasmic localization of MEK1. Panel e shows no primary antibody. The images were captured at 20X magnification.