710859

antibody from Invitrogen Antibodies

Targeting: APC DP2, DP2.5, DP3, PPP1R46

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

710859

Provider

Invitrogen Antibodies

Product name

Protein APC Recombinant Polyclonal Antibody (1HCLC)

Antibody type

Polyclonal

Antigen

Other

Description

This antibody is predicted to react with Monkey, Cat, Pig and Bovine.

Antibody clone number

1HCLC

Concentration

0.5 mg/mL

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis was performed on fixed and permeabilized MCF-7 cells for detection of endogenous APC using Anti-APC Recombinant Rabbit Polyclonal Antibody (Product # 710859, 2 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Panel a) shows representative cells that were stained for detection and localization of APC protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Panel c) represents cytoskeletal F-actin staining using Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d) is a composite image of Panels a, b and c clearly demonstrating cytoplasmic localization of APC. 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

Immunofluorescence analysis was performed on fixed and permeabilized MCF-7 cells for detection of endogenous APC using Anti-APC Recombinant Rabbit Polyclonal Antibody (Product # 710859, 2 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Panel a) shows representative cells that were stained for detection and localization of APC protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Panel c) represents cytoskeletal F-actin staining using Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d) is a composite image of Panels a, b and c clearly demonstrating cytoplasmic localization of APC. 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

Immunofluorescence analysis of Protein APC was performed using 70% confluent log phase HT29 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with Protein APC Recombinant Polyclonal Antibody (1HCLC) (Product # 710859) at 1:100 in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32766), (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). Panel d represents the merged image showing plasma membrane localization for Protein APC in HeLa. Panel e represents control HeLa 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

Knockdown of Protein APC was achieved by transfecting HT29 cells with Protein APC specific siRNA (Silencer® select Product # S1435, S1433). Immunofluorescence analysis was performed on untransfected HT29 cells (panel a,d), transfected with non-specific scrambled siRNA (panels b,e) and transfected with Protein APC specific siRNA (panel c,f). Cells were fixed, permeabilized, and labelled with Pericentrin Rabbit Polyclonal Antibody (Product # PA5-54109, 4 µg/mL), followed by Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjµgate (Product # A27034, 1:2000). Nuclei (blue) were stained using ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962), and Rhodamine Phalloidin (Product # R415, 1:300) was used for cytoskeletal F-actin (red) staining. Reduction of specific signal was observed upon siRNA mediated knockdown (panel c,f) confirming specificity of the antibody to Protein APC (green). The images were captured at 60X magnification.