Anti-FADD antibody [EPR5030] - BSA and Azide free

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-FADD antibody [EPR5030] - BSA and Azide free (AB229444)

This data was developed using ab124812, the same antibody clone in a different buffer formulation. Intracellular Flow Cytometry analysis of NIH/3T3 (Mouse embryonic fibroblast) cells labelling FADD with purified ab124812 at 1/40 dilution (10 �g/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). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FADD antibody [EPR5030] - BSA and Azide free (AB229444)

This data was developed using ab124812, the same antibody clone in a different buffer formulation.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue sections labeling FADD with purified ab124812 at 1/150 dilution (2.59 μg/mL). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1/0 dilution. PBS instead of the primary antibody was used as the negative control.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-FADD antibody [EPR5030] - BSA and Azide free (AB229444)

This data was developed using ab124812, the same antibody clone in a different buffer formulation.

Immunocytochemistry analysis of NIH/3T3 (Mouse embryonic fibroblast) cells labeling FADD with purified ab124812 at 1/50 dilution (7.8 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 (2.5 μg/mL). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/1000 (2 μg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• IP

Lab

Immunoprecipitation - Anti-FADD antibody [EPR5030] - BSA and Azide free (AB229444)

FADD was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 10 μg with ab124812 at 1/30 dilution (2 µg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab124812 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution. Lane 1 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 10 μg. Lane 2 : NIH/3T3 whole cell lysate. Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab124812 in NIH/3T3 whole cell lysate. Blocking and dilution buffer and concentration : 5% NFDM/TBST. Observed MW : 28 kDa. Exposure time : 41 secs. This data was developed using ab124812, the same antibody clone in a different buffer formulation.

• IP

Lab

Immunoprecipitation - Anti-FADD antibody [EPR5030] - BSA and Azide free (AB229444)

FADD was immunoprecipitated from 0.35 mg RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate 10 μg with ab124812 at 1/30 dilution (2 µg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab124812 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution. Lane 1 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate 10 μg. Lane 2 : RAW264.7 whole cell lysate. Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab124812 in RAW264.7 whole cell lysate. Blocking and dilution buffer and concentration : 5% NFDM/TBST. Observed MW : 28 kDa. Exposure time : 3 minutes. This data was developed using ab124812, the same antibody clone in a different buffer formulation.

• WB

Unknown

Western blot - Anti-FADD antibody [EPR5030] - BSA and Azide free (AB229444)

All lanes:

Western blot - Anti-FADD antibody [EPR5030] (<a href='/en-us/products/primary-antibodies/fadd-antibody-epr5030-ab124812'>ab124812</a>) at 1/1000 dilution

All lanes:

NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 23 kDa

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