MA1-19178

antibody from Invitrogen Antibodies

Targeting: AFP FETA, HPAFP

ELISA

Immunoprecipitation

Antibody data

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Validation data

Product number: MA1-19178 - Provider product page
Provider: Invitrogen Antibodies
Product name: AFP Monoclonal Antibody (AFP-01)
Antibody type: Monoclonal
Antigen: Purifed from natural sources
Description: This antibody reacts with human alpha-Fetoprotein (AFP), a 70 kDa oncofetal antigen. AFP is a major fetal plasma protein, but is not present in healthy adult tissues. Elevated AFP concentrations in adult plasma may be an early marker of hepatocellular carcinoma or teratoblastoma, while high concentrations in amniotic fluid may indicate severe congenital defects of a fetus. This antibody has successfully been paired as the coating antibody in a sandwich ELISA with detection antibody MA1-19178 (clone AFP-01). Immunoprecipitation: Interaction of the antibody AFP-01 with AFP is dependent on the presence of calcium ions (strongly inhibited by chelating agents). Such characteristics of the antibody can be exploited for immunoaffinity purification of APF under mild elution conditions; Western Blot: non-reducing conditions.
Reactivity: Human
Host: Mouse
Isotype: IgG
Antibody clone number: AFP-01
Vial size: 100 µg
Concentration: 1 mg/mL
Storage: 4° C, do not freeze

AFP protein structure - MA1-19178 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of alpha Fetoprotein was performed on HepG2 whole cell lysates. Cell lysates were mixed with reducing (Lane 1) or non-reducing (Lane 2) Laemmli SDS-PAGE sample buffer and 5 µL of lysate, corresponding to 6.25 x 10^5 HepG2 cells, was loaded per well on a 10% polyacrylamide gel. Proteins were transferred to a membrane and the membrane was blocked with 5% milk overnight at 4C. The membrane was probed with an alpha Fetoprotein monoclonal antibody (Product # MA1-19178) at a dilution of 1:500-1:2000 for 1 hour at RT, followed by a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:1000 for 1 hour at RT. Detection was performed using a chemiluminescent substrate

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on Conditioned Media of Hep G2 (Lane 1) and HeLa (Lane 2). The blot was probed with anti-AFP Mouse Monoclonal Antibody (Product # MA1-19178, 1:1000 dilution) and detected by chemiluminescence using Goat anti Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/mL, 1:4000 dilution). A 68 kDa band corresponding to AFP was detected in Hep G2 conditioned media but not in HeLa conditioned media which is negative for AFP expression.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of alpha Fetoprotein was performed on HepG2 whole cell lysates. Cell lysates were mixed with reducing (Lane 1) or non-reducing (Lane 2) Laemmli SDS-PAGE sample buffer and 5 µL of lysate, corresponding to 6.25 x 10^5 HepG2 cells, was loaded per well on a 10% polyacrylamide gel. Proteins were transferred to a membrane and the membrane was blocked with 5% milk overnight at 4C. The membrane was probed with an alpha Fetoprotein monoclonal antibody (Product # MA1-19178) at a dilution of 1:500-1:2000 for 1 hour at RT, followed by a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:1000 for 1 hour at RT. Detection was performed using a chemiluminescent substrate

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blotting analysis of human alpha-fetoprotein using mouse Monoclonal antibody AFP-01 (Product # MA1-19178) on lysates of HepG2 cell line and Jurkat cell line (negative control) under reducing and non-reducing conditions. Nitrocellulose membrane was probed with 2µg/mL of mouse monoclonal antibody anti-alpha-fetoprotein followed by IRDye800-conjugated anti-mouse secondary antibody. Alpha-fetoprotein was detected at approximately 60kDa.