480028

antibody from Invitrogen Antibodies

Targeting: PLK1 PLK

Antibody data

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Validation data

PLK1 protein structure - 480028 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot of rat synaptic membrane showing specific immunolabeling of the ~66 kD PLK protein phosphorylated at Thr210 (control). The phosphospecificity of this labeling is shown in the second lane (lambda-phosphatase: L-Ptase). The blot is identical to the control except that it was incubated in L-Ptase (1200 units for 30 min) before being exposed to the phospho-Thr210 PLK antibody. The immunolabeling is completely eliminated by treatment with L-Ptase.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-Phospho-PLK1 (Thr210) Polyclonal Antibody (Product # 480028) and a 68kDa band corresponding to phosphorylated Serine/threonine-protein kinase PLK1 was observed in HeLa treated with Calyculin A. A non specific band around 22kDa was also observed. Whole cell extracts (30 µg lysate) of HeLa (Lane 1) and HeLa treated with Calyculin A (25nM for 30 min) (Lane 2) were electrophoresed using NuPAGE™ 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 the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot of PLK1 in rat synaptic membrane showing specific immunolabeling of a ~66 kDa band corresponding to Phospho-PLK1 (Thr210) polyclonal antibody (Product # 480028) in the first lane (-). Phosphospecificity is shown in the second lane (+) where the immunolabeling is completely eliminated by blot treatment with lambda phosphatase (1,200 units for 30 minutes).