- MA5-33127 - Invitrogen Antibodies CDC25C antibody

No submitted references: Submit reference

No comments: Submit comment

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of Cdc25C using a Cdc25C Monoclonal antibody (Product # MA5-33127) at a concentration of 1.65 µg/mL. Positive WB detected in: Hela whole cell lysate, K562 whole cell lysate, Raji whole cell lysate, 293 whole cell lysate. A secondary Goat polyclonal antibody to rabbit IgG was applied at a 1:50,000 dilution. Observed band size: 60 kDa.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of precipitated Cdc25C from HEK293 whole cell lysate using a Cdc25C monoclonal antibody (Product # MA5-33127). An HRP-conjugated Protein G antibody was used as the secondary antibody (1:2000). Lane 1: Rabbit control IgG. Lane 2: HEK293 whole cell lysate (500µg). Lane 3: HEK293 whole cell lysate (20µg).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of Cdc25C using a Cdc25C Monoclonal antibody (Product # MA5-33127) at a concentration of 1.65 µg/mL. Positive WB detected in: Hela whole cell lysate, K562 whole cell lysate, Raji whole cell lysate, 293 whole cell lysate. A secondary Goat polyclonal antibody to rabbit IgG was applied at a 1:50,000 dilution. Observed band size: 60 kDa.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot was performed using Anti-Cdc25C Recombinant Rabbit Monoclonal Antibody (Product # MA5-33127) and a 50, 53 kDa band corresponding to M-phase inducer phosphatase 3 was observed across the cell lines tested. Nuclear enriched extracts (30 µg lysate) of HCT 116 (Lane 1), HCT 116 treated with 5 nm Actinomycin D for 48 hours (Lane 2), HT-29 (Lane 3), HT-29 treated with 0.1 UM Nocodazole for 4 hours (Lane 4) and K-562 (Lane 5) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # LC2001) 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: Immunofluorescent analysis of Cdc25C in Hela cells using a Cdc25C monoclonal antibody (Product # MA5-33127) at a dilution of 1:55. The cells were fixed in 4% formaldehyde, permeabilized using 0.2% Triton X-100 and blocked in 10% normal Goat Serum. The cells were then incubated with the antibody overnight at 4°C. The secondary antibody was Alexa Fluor 488-congugated Goat Anti-Rabbit IgG (H+L). Cells were counter-stained with DAPI.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of M-phase inducer phosphatase 3 was performed using 70% confluent log phase HCT 116 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 Cdc25C Recombinant Rabbit Monoclonal Antibody (Product # MA5-33127) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000 dilution), 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 dilution). Panel d represents the merged image showing nuclear speckle localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemical analysis of Cdc25C in paraffin embedded human glioma cancer using a Cdc25C monoclonal antibody (Product # MA5-33127) at a dilution of 1:165. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a biotinylated secondary antibody and visualized using an HRP conjugated SP system.

MA5-33127