EM1902-25

DAP Kinase 1 Mouse Monoclonal Antibody [12B1]

cat.: EM1902-25

Product Type:	Mouse monoclonal IgG1, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IHC-P, IF-Cell
Clonality:	Monoclonal
Clone number:	12B1

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	2ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 160 kDa
Isotype:	IgG1
Immunogen:	Recombinant protein within Human DAPK1 aa 447-683 / 1,430.
Positive control:	Human skin tissue lysate, human skeletal muscle tissue lysate, MCF-7, rat lung tissue, human lung cancer tissue, mouse liver tissue, LOVO.
Subcellular location:	Cytoplasm, Cytoskeleton.
Recommended Dilutions: WB IHC-P IF-Cell	1:500-1:2,000 1:50-1:200 1:50-1:100
Uniprot #:	SwissProt: P53355 Human \| Q80YE7 Mouse
Alternative names:	DAK1 DAP K1 DAP kinase 1 DAPK 1 DAPK DAPK1 DAPK1_HUMAN Death Associated Protein Kinase 1 Death-associated protein kinase 1 DKFZp781I035

Images

	Fig1: Western blot analysis of DAP Kinase 1 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (EM1902-25, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature. Positive control: Lane 1: Human skin tissue lysate Lane 2: Human skeletal muscle tissue lysate
	Fig2: ICC staining of DAP Kinase 1 in MCF-7 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (EM1902-25, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Mouse IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).
	Fig3: Immunohistochemical analysis of paraffin-embedded rat lung tissue using anti-DAP Kinase 1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (EM1902-25, 1/100) 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 lung cancer tissue using anti-DAP Kinase 1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (EM1902-25, 1/100) 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 liver tissue using anti-DAP Kinase 1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (EM1902-25, 1/100) 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: Immunocytochemistry analysis of LOVO cells labeling DAP Kinase 1 with Mouse anti-DAP Kinase 1 antibody (EM1902-25) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 30 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and then blocked with 2% BSA for 30 minutes at room temperature. Cells were then incubated with Mouse anti-DAP Kinase 1 antibody (EM1902-25) at 1/50 dilution in 2% BSA overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

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