MA1-35436

antibody from Invitrogen Antibodies

Targeting: AFP FETA, HPAFP

Western blot (Data presented on provider website)

ELISA (Recommended by provider)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Recommended by provider)

Antibody Data

Product number

MA1-35436

Provider

Invitrogen Antibodies

Product name

AFP Monoclonal Antibody (C2)

Antibody type

Monoclonal

Antigen

Other

Description

C2 has been characterized in the ISOBM TD-2 workshop and assigned by K. Nustad to group E of a cluster of 6 major epitopes of human alpha fetoprotein. Human alpha fetoprotein is an oncofetal protein of 70 kDa. It is expressed in fetal liver and is normally absent in health adult tissues. It is positive on all yolk sac tumors, on some other germ cell tumors and on hepatocellular carcinomas.

Reactivity

Human, Mouse

Host

Mouse

Isotype

IgG

Antibody clone number

C2

Vial size

100 μg

Concentration

100 μg/mL

Storage

4°C

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of AFP was performed using 70% confluent log phase Hep G2 cells 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 1 hour at room temperature. The cells were labeled with AFP Monoclonal Antibody (C2) (Product # MA1-35436) at 1:100 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, 1:2,000), 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 and golgi staining of AFP in Hep G2 cells. Panel e represents MCF7 with no signal. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of AFP was performed using 70% confluent log phase Hep G2 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 1 hour at room temperature. The cells were labeled with AFP Monoclonal Antibody (C2) (Product # MA1-35436) at 1:100 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, 1:2,000), 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 and golgi staining of AFP in Hep G2 cells. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.