711092

antibody from Invitrogen Antibodies

Targeting: FASN FAS, SDR27X1

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

711092

Provider

Invitrogen Antibodies

Product name

FASN Recombinant Polyclonal Antibody (10HCLC)

Antibody type

Polyclonal

Antigen

Other

Reactivity

Human

Host

Rabbit

Isotype

IgG

Antibody clone number

10HCLC

Vial size

100 µg

Concentration

0.5 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on Membrane enriched extracts (30 µg lysate) of Hep G2 (Lane 1), MCF-7 (Lane 2), Caco-2 (Lane 3), A549 (Lane 4), HeLa (Lane 5), NIH/3T3 (Lane 6), NTERA-2 (Lane 7) and PC-3 (Lane 8). The blots were probed with Anti-Fatty acid synthase Recombinant Rabbit Polyclonal Antibody (Product # 711092, 1-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 273 kDa band corresponding to Fatty acid synthase protein was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex®NuPAGE®4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane using wet transfer method. The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

For immunofluorescence analysis, Hep G2 cells were fixed and permeabilized for detection of endogenous Fatty acid synthase using Anti- Fatty acid synthase Recombinant Rabbit Polyclonal Antibody (Product # 711092, 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 Fatty acid synthase 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 Fatty acid synthase. Panel e) represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.