- PA5-18509 - Invitrogen Antibodies CSNK1D antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot staining of HEK293 cell lysate using Product # PA5-18509 at a concentration of 0.3 µg/mL, the primary antibody incubation was 1 hour and the detection method was chemiluminescence.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot staining of HEK293 cell lysate using Product # PA5-18509 at a concentration of 0.3 µg/mL, the primary antibody incubation was 1 hour and the detection method was chemiluminescence.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of CK1 delta was achieved by transfecting HeLa with CK1 delta specific siRNAs (Silencer® select Product # s3627 and s3628). Western blot analysis (Fig. a) was performed using Whole cell extracts from the CK1 delta knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with CK1 delta Polyclonal Antibody (Product # PA5-18509, 0.75 µg/ml) and Rabbit anti-Goat IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27014, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to CK1 delta.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using Anti-CK1 delta Polyclonal Antibody (Product # PA5-18509) and a 49 kDa band corresponding to CK1 delta was observed across cell lines tested. Whole cell extracts (30 µg lysate) of NIH/3T3 (Lane 1), MCF7 (Lane 2), HEK-293 (Lane 3) and HeLa (Lane 4) 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 (0.75 µg/mL) and detected by chemiluminescence with Rabbit anti-Goat IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27014, 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)
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Experimental details: Immunocytochemical analysis of CK1 delta in U251 cells using a CK1 delta polyclonal antibody (Product #PA5-18509). U251 cells were permeabilized with 0.15% Triton. Cells were incubated with 10 µg/mL of primary antibody for one hour followed by an Alexa Fluor 488 secondary antibody at a concentration of 2 µg/mL. Nuclear staining can be seen as shown above. The nuclear stain is DAPI (blue). Negative control: Unimmunized goat IgG (10 µg/mL) followed by an Alexa Fluor 488 secondary antibody (2 µg/mL).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of CK1 delta was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with CK1 delta Polyclonal Antibody (Product # PA5-18509) at 5 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Goat IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32814), (1:2000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear, cytoplasmic (golgi, ER) and cytoskeletal localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

PA5-18509