PA5-18672

antibody from Invitrogen Antibodies

Targeting: PNPLA2 ATGL, desnutrin, FP17548, iPLA2zeta, TTS-2.2

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-18672

Provider

Invitrogen Antibodies

Product name

ATGL Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

This antibody is predicted to react with bovine, canine, mouse, porcine and rat based on sequence homology.

Reactivity

Human

Host

Goat

Isotype

IgG

Vial size

100 µg

Concentration

0.5 mg/mL

Storage

-20° C, Avoid Freeze/Thaw Cycles

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot staining of Human Adipose lysate using Product # PA5-18672 at a concentration of 0.5 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 was performed using ATGL Polyclonal Antibody (Product # PA5-18672) and a 55 kDa band corresponding to ATGL was observed across cell lines and tissues tested. Whole cell extracts (30 µg lysate) of Raw 264.7 (Lane 1), NIH: OVCAR-3 (Lane 2), NIH/3T3 (Lane 3), HEK-293 (Lane 4), Mouse adipose (Lane 5), Mouse skeletal muscle (Lane 6), Rat adipose (Lane 7), Rat skeletal muscle (Lane 8), 3T3-L1 (Lane 9), 3T3-L1 differentiated to adipocytes (Lane 10) were electrophoresed using NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX). 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 (0.5µg/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).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of ATGL was performed using 70% confluent log phase 3T3-L1 and 3T3-L1 differentiated to adipocytes. 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 ATGL Polyclonal Antibody (Product # PA5-18672) at 5 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant 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 localization to vacuolar structure (fat droplets) in cytoplasm. Panel e shows 3T3-L1 cells with no expression of ATGL. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical staining of paraffin embedded of Human Heart using Product # PA5-18672 at a concentration of 2.5 µg/mL. The tissue was processed by steamed antigen retrieval with citrate buffer pH 6 and stained with AP.