PA5-66752

antibody from Invitrogen Antibodies

Targeting: AP1B1 ADTB1, AP105A, BAM22, CLAPB2

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-66752

Provider

Invitrogen Antibodies

Product name

beta-1 Adaptin Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant full-length protein

Description

Immunogen sequence: PPSAFVEGGRG VVHKSLPPRT ASSESAESPE TAPTGAPP

Concentration

0.7 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of beta-1 Adaptin in human cell line RT-4, human cell line U-251 MG, human plasma, human liver tissue and human tonsil tissue. Samples were probed using a beta-1 Adaptin Polyclonal Antibody (Product # PA5-66752).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using anti-beta-1 Adaptin Polyclonal Antibody (Product # PA5-66752) and a 110kDa band corresponding to beta-1 Adaptin was observed across all the cell lines and tissue lysates tested except Mouse Skeletal Muscle and Rat Skeletal Muscle which are reported negative for beta-1 Adaptin expression. An additional band which corresponds to an isoform of beta-1 Adaptin was detected at 65KDa in MCF7, MOLT-4, HepG2, Jurkat and LNCaP. Whole cell extracts (30 µg lysate) of HEK-293 (Lane 1), MCF7 (Lane 2), MOLT-4 (Lane 3), Hep G2 (Lane 4), Jurkat (Lane 5), LNCaP (Lane 6), A549 (Lane 7), tissue extracts of Mouse Skeletal Muscle (Lane 8), Rat Skeletal Muscle (Lane 9), Mouse Brain (Lane 10) and Rat Brain (Lane 11) were electrophoresed using NuPAGE® 10 % Bis-Tris gel (Product # NP0302BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blots were probed with the primary antibody (1:200 dilution) and detected by chemiluminescence Goat Anti-Rabbit IgG Secondary Antibody, HRP conjugate (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

Immunofluorescent staining of beta-1 Adaptin in human cell line HEK 293 shows localization to cytosol and the Golgi apparatus. Samples were probed using a beta-1 Adaptin Polyclonal Antibody (Product # PA5-66752).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of beta-1 Adaptin was performed using 70% confluent log phase HEK-293 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with beta-1 Adaptin Polyclonal Antibody (Product # PA5-66752) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 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, golgi and vesicular localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification. .