PA1-4652

antibody from Invitrogen Antibodies

Targeting: DDC AADC

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA1-4652

Provider

Invitrogen Antibodies

Product name

DOPA Decarboxylase Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

This antibody is specific for the ~55 kDa DDC protein in Western blots of rat adrenal medulla.

Reactivity

Human, Rat, Bovine, Canine, Guinea Pig, Rabbit

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Storage

-20° C, Avoid Freeze/Thaw Cycles

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot of 5 µg of bovine adrenal medulla lysate showing specific immunolabeling of the ~55 kDa DOPA Decarboxylase protein.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot of DOPA Decarboxylase in bovine adrenal medulla lysate (5 µg). This shows specific immunolabeling of the ~55 kDa DOPA Decarboxylase polyclonal antibody (Product # PA1-4652).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-DOPA Decarboxylase Polyclonal Antibody (Product # PA1-4652) and ~53 kDa band corresponding to DOPA Decarboxylase was observed in IMR-32, SH-SY5Y and Caco-2, but not in HEL.92.1.7 and SK-O-V3 which are reported to be negative for DOPA Decarboxylase expression. Whole cell lysate (40ug lysate) of IMR-32 (Lane 1), SH-SY5Y (Lane 2), Caco-2 (Lane 3), HEL 92.1.7 (Lane 4) and SK-O-V3 (Lane 5) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The bot 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).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of DOPA Decarboxylase was performed using 70% confluent log phase SH-SY5Y and SK-O-V3 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 10 minutes at room temperature. The cells were labeled with DOPA Decarboxylase Polyclonal Antibody (Product # PA1-4652) 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 SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic and nuclear localization for DOPA Decarboxylase in SH-SY5Y cells. Panel e represents merged image of SK-O-V3, which is reported to be low expressing for the same. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.