Anti-alpha 1 Fetoprotein antibody [EPAFP61] - Low endotoxin, Azide free

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-alpha 1 Fetoprotein antibody [EPAFP61] - Low endotoxin, Azide free (AB215379)

This data was developed using ab133617, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling alpha 1 Fetoprotein with Purified ab133617 at 1/250 dilution (0.52 µg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 dilution (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 dilution (2 µg/mL). DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha 1 Fetoprotein antibody [EPAFP61] - Low endotoxin, Azide free (AB215379)

This data was developed using ab133617, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver cancer tissue sections labeling alpha 1 Fetoprotein with Purified ab133617 at 1/100 dilution (1.3 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 1 (pH 6.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-alpha 1 Fetoprotein antibody [EPAFP61] - Low endotoxin, Azide free (AB215379)

This data was developed using ab133617, the same antibody clone in a different buffer formulation. Intracellular Flow Cytometry analysis of HepG2 Human hepatocellular carcinoma epithelial cell cells labeling alpha 1 Fetoprotein with purified ab133617 at 1/100 dilution (10 μg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cells without incubation with primary antibody and secondary antibody (Blue).

• IP

Lab

Immunoprecipitation - Anti-alpha 1 Fetoprotein antibody [EPAFP61] - Low endotoxin, Azide free (AB215379)

This data was developed using ab133617, the same antibody clone in a different buffer formulation.
Purified ab133617 at 1/20 dilution (0.5μg) immunoprecipitating alpha 1 Fetoprotein in HepG2 whole cell lysate.
Lane 1 (input) : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate 10μg
Lane 2 (+) : ab133617 + HepG2 whole cell lysate.
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab133617 in HepG2 whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/5000 dilution) was used for Western blotting.
Blocking Buffer and concentration : 5% NFDM/TBST.
Diluting buffer and concentration : 5% NFDM/TBST.
Observed band size : 70 kDa
Lower bands could be caspase-dependent cleavage fragments. (PMID : 31811908)

All lanes:

Immunoprecipitation - Anti-alpha 1 Fetoprotein antibody [EPAFP61] (<a href='/en-us/products/primary-antibodies/alpha-1-fetoprotein-antibody-epafp61-ab133617'>ab133617</a>)

Predicted band size: 68 kDa

false

• WB

Unknown

Western blot - Anti-alpha 1 Fetoprotein antibody [EPAFP61] - Low endotoxin, Azide free (AB215379)

This data was developed using ab133617, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-alpha 1 Fetoprotein antibody [EPAFP61] (<a href='/en-us/products/primary-antibodies/alpha-1-fetoprotein-antibody-epafp61-ab133617'>ab133617</a>) at 1/10000 dilution

All lanes:

HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate at 15 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 68 kDa

false