437100

antibody from Invitrogen Antibodies

Targeting: ACACA ACAC, ACC, ACC1

Western blot (Supportive data in Antibodypedia)

ELISA (Recommended by provider)

Immunoprecipitation (Recommended by provider)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

437100

Provider

Invitrogen Antibodies

Product name

Acetyl-CoA Carboxylase Monoclonal Antibody (143)

Antibody type

Monoclonal

Antigen

Recombinant full-length protein

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

143

Vial size

100 µg

Concentration

0.5 mg/mL

Storage

-20°C

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of ACC1 was performed using membrane enriched extracts (30 µg lysate) of A549 (Lane 1), K-562 (Lane 2), HeLa (Lane 3), PC-3 (Lane 4), HEK-293 (Lane 5), Caco-2 (Lane 6) and Hep G2 (Lane 7). The blots were probed with Anti-ACC1 Mouse monoclonal Antibody (Product # 437100, 1-3 µg/mL) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.4 µg/mL, 1:2500 dilution). A 265 kDa band corresponding to ACC1 were observed across the cell lines tested. Along with the desired bands, non-specific bands at 120 kDa were observed across cell lines, except PC-3. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and HiMark™ Pre-stained Protein Standard (Product # LC5699). Resolved proteins were then transferred onto a nitrocellulose membrane with overnight wet transfer System. 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).

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry analysis of ACC1 was performed using Hep G2 cells. Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Triton™ X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with ACC1 Mouse Monoclonal Antibody (437100, red histogram) or with mouse isotype control (pink histogram) at 3-5 ug/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor® 488 Rabbit Anti-Mouse Secondary Antibody (A11059) at a dilution of 1:400 for 30 minutes at room temperature. The representative 10,000 cells were acquired and analyzed for each sample using an Attune® Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control..