MA5-14665

antibody from Invitrogen Antibodies

Targeting: AFP FETA, HPAFP

Western blot (Data presented on provider website)

ELISA (Recommended by provider)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunoprecipitation (Recommended by provider)

Radioimmunoassay (Recommended by provider)

Antibody Data

Product number

MA5-14665

Provider

Invitrogen Antibodies

Product name

AFP Monoclonal Antibody (F1-6P2A8-P2B9A9)

Antibody type

Monoclonal

Antigen

Purifed from natural sources

Description

MA5-14665 can be used for immunofluorescent analysis of AFP in the endoderm derived from human embryonic stem cells. By Western blot, MA5-14665 detects endogenous AFP protein in the early hepatocyte-like cells derived from human embryonic stem cells. Product MA514665 is a smaller package size of MIA1301 (formerly sold as a Seradyn product).

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

F1-6P2A8-P2B9A9

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

Immunofluorescence analysis of alpha-fetoproprotein (AFP) (green) in the endoderm derived from human ES cells. Embryoid bodies (EBs) were generated from the H9 embryonic stem cell line (WiCell Research Institute, WA09) using Gibco® KnockOut™ Serum Replacement. After four days in suspension culture, EBs were plated on Geltrex™-coated tissue culture-treated polystyrene plates and continuously cultured for 21 days. EB cultures were then fixed and permeabilized according to the 3-Germ Layer Immunocytochemistry Kit (Product # A25538) and stained with anti-alpha-fetoprotein (Product # MA5-14665, 1:200 dilution, 5 uL/mL final) at 4°C overnight. Secondary staining was completed using Alexa Fluor™ 488-conjugated anti-mouse IgG (Product # A-11001) and DAPI (Product # D1306) for nuclear DNA (blue) for 1 h at room temperature. Stained wells were imaged at 40X using the EVOS® FL Auto Imaging System.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of AFP was performed using 70% confluent log phase Hep G2 and MDA-MB-231 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 Mouse Monoclonal Antibody (Product # MA5-14665) at 5 µg/mL in 0.1% BSA and incubated overnight at 4 degree and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 (Product # A28175) at a dilution of 1:2000 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 composite image showing cytoplasmic and golgi staining of AFP in Hep G2. Panel e represents the merged image of MDA-MB-231 cells which do not have AFP expression. 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 and MDA-MB-231 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 Mouse Monoclonal Antibody (Product # MA5-14665) at 5 µg/mL in 0.1% BSA and incubated overnight at 4 degree and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 (Product # A28175) at a dilution of 1:2000 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 composite image showing cytoplasmic and golgi staining of AFP in Hep G2. Panel e represents the merged image of MDA-MB-231 cells which do not have AFP expression. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification..