ET1612-2

MEK3 Recombinant Rabbit Monoclonal Antibody [SD20-93]

cat.: ET1612-2

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IF-Tissue, IHC-P, IP, FC
Clonality:	Monoclonal
Clone number:	SD20-93

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 39 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within human MEK3 aa 101-347/347.
Positive control:	Hela cell lysates, Hela, human liver tissue, human breast carcinoma tissue, mouse spleen tissue.
Subcellular location:	Cytosol, nucleoplasm, membrane.
Recommended Dilutions: WB IF-Cell IF-Tissue IHC-P FC	1:1,000-1:5,000 1:50-1:200 1:50-1:200 1:50-1:200 1:50-1:100
Uniprot #:	SwissProt: P46734 Human \| O09110 Mouse Unigene: 100064 Rat
Alternative names:	AW212142 dual specificity mitogen activated protein kinase kinase 3 Dual specificity mitogen-activated protein kinase kinase 3 MAP kinase kinase 3 map2k3 MAPK ERK kinase 3 MAPK/ERK kinase 3 MAPKK 3 MAPKK3 MEK 3 MEK3 Mitogen activated protein kinase kinase 3 MKK 3 MKK3 mMKK3b MP2K3_HUMAN PRKMK 3 PRKMK3 protein kinase, mitogen-activated, kinase 3 SAPK kinase 2 SAPKK 2 SAPKK2 Stress activated protein kinase kinase 2

Images

	Fig1: Western blot analysis of MEK3 on Hela cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1612-2, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature. Predicted band size: 39 kDa Observed band size: 39 kDa
	Fig2: ICC staining of MEK3 in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1612-2, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
	Fig3: Immunohistochemical analysis of paraffin-embedded human liver tissue using anti-MEK3 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1612-2, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig4: Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-MEK3 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1612-2, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

	Fig5: Immunohistochemical analysis of paraffin-embedded mouse spleen tissue using anti-MEK3 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1612-2, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig6: Flow cytometric analysis of MEK3 was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1612-2, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
	Fig7: Western blot analysis of MEK3 on different lysates with Rabbit anti-MEK3 antibody (ET1612-2) at 1/2,000 dilution. Lane 1: A549-si NT cell lysate Lane 2: A549-si MEK3 cell lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 39 kDa Observed band size: 39 kDa Exposure time: 30seconds; ECL: K1801; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1612-2) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.