MA5-14736

antibody from Invitrogen Antibodies

Targeting: FTL MGC71996, NBIA3

ELISA (Data presented on provider website)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunoprecipitation (Recommended by provider)

Radioimmunoassay (Recommended by provider)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-14736

Provider

Invitrogen Antibodies

Product name

Ferritin Monoclonal Antibody (513C10)

Antibody type

Monoclonal

Antigen

Other

Description

MA5-14736 detects Ferritin in ELISA, IP, IF, and RIA applications and shows reactivity to human samples. The MA5-14736 Immunogen is human liver ferritin. MA5-14736 detects ferritin which has a predicted molecular weight of approximately 19kDa. Product MA514736 is a smaller package size of MIF2504 (formerly sold as a Seradyn product).

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

513C10

Vial size

100 µg

Concentration

1 mg/mL

Storage

Maintain refrigerated at 2-8°C for up to 6 months. For long term storage store at -20°C

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of Ferritin (FTL, green) in HepG2 cells and MCF7 cells. The cells were fixed with formalin for 15 minutes, permeabilized with 0.1% Triton X-100 in TBS for 10 minutes, and blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were stained with a Ferritin monoclonal antibody (Product # MA5-14736), at a dilution of 1:200 for at least 1 hour at room temperature, and then incubated with a DyLight 488 goat anti-mouse IgG secondary antibody (Product # 35503) at a dilution of 1:500 for 30 minutes at room temperature (both panels, green). F-Actin (both panels, red) was stained with DyLight 554 Phalloidin (Product # 21834) and nuclei (both panels, blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ToxInsight at 20X magnification.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Ferritin was performed using 70% confluent log phase Hep G2 and MCF7 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with Ferritin Monoclonal Antibody (513C10) (Product # MA5-14736) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32766), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e represents MCF7 cells with reduced expression. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Direct ELISA analysis of Ferritin was performed by coating wells of a 96-well plate with 100 µL per well of Human Ferritin or BSA diluted in carbonate/bicarbonate buffer (Product # 28382), starting at a concentration of 1 µg/mL and serially diluting 2-fold to a concentration of 2 ng/mL, overnight at 4C. Wells of the plate were washed, blocked with StartingBlock blocking buffer (Product # 37538), and incubated with 100 µL per well of a mouse anti-ferritin monoclonal antibody (Product # MA5-14736) at a concentration of 1 µg/mL for 1 hour at room temperature. The plate was washed, then incubated with 100 µL per well of an HRP-conjugated goat anti-mouse IgG secondary antibody (Product # 31430) at a dilution of 1:25000 for 30 minutes at room temperature. Detection was performed using 1-Step Ultra TMB substrate (Product # 34028) for 10 minutes at room temperature in the dark. The reaction was stopped with 0.16M sulfuric acid, and absorbances were read on a spectrophotometer at 450-550 nm.