Anti-Fas 抗体 [EPR5700]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fas antibody [EPR5700] (AB133619)

Immunohistochemical analysis of paraffin embedded Human tonsil tissue labelling CD95 with ab133619 antibody at a dilution of 1/250.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Fas antibody [EPR5700] (AB133619)

Immunocytochemistry analysis of Raji (Human Burkitt's lymphoma B lymphocyte) labeling Fas with purified ab133619 at 1/50 dilution. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/1000 (2 µg/ml) was used as the secondary antibody. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.31 µg/ml) was used as counterstain. Nuclei were stained blue with DAPI.
Negative control : PBS instead of the primary antibody.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fas antibody [EPR5700] (AB133619)

Anti-Fas antibody [EPR5700] (ab133619)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling Fas with ab133619 at a dilution of 1 : 500. Heat mediated antigen retrieval was performed using AR9 antigen retrieval solution, and microwave treatment for 15 min at 20% power. Anti-Rabbit/Mouse HRP polymer (PerkinElmer Opal Polymer HRP Ms Plus Rb) was used as secondary antibody. Opal tyramide amplification was performed using Opal 520 fluorophore. Counterstained with DAPI stain. Image scanned with Vectra 3.0 and analyzed via Phenochart software.
This image was courteously provided by Dr. Houssein Abdul Sater, Georgia Cancer Center.

• WB

Lab

Western blot - Anti-Fas antibody [EPR5700] (AB133619)

Lanes 1 - 4 : Merged signal (red and green). Green - ab133619 observed at 42 kDa. Red - loading control, ab8245, observed at 38 kDa.

ab133619 was shown to specifically react with Fas when Fas knockout samples were used. Wild-type and Fas knockout samples were subjected to SDS-PAGE. ab133619 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10 000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-Fas antibody [EPR5700] (ab133619) at 1/1000 dilution

Lane 1:

Wild-type HAP1 cell lysate at 20 µg

Lane 2:

Fas knockout HAP1 cell lysate at 20 µg

Lane 3:

Raji cell lysate at 20 µg

Lane 4:

Ramos cell lysate at 20 µg

Predicted band size: 37 kDa

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• WB

Lab

Western blot - Anti-Fas antibody [EPR5700] (AB133619)

Lanes 1-2 : Merged signal (red and green). Green - ab133619 observed at 37 kDa. Red - loading control ab8245 observed at 37 kDa.

ab133619 Anti-Fas antibody [EPR5700] was shown to specifically react with Fas in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265260 (knockout cell lysate ab256911) was used. Wild-type and Fas knockout samples were subjected to SDS-PAGE. ab133619 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Fas antibody [EPR5700] (ab133619) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human FAS knockout HeLa cell line (<a href='/products/cell-lines/human-fas-knockout-hela-cell-line-ab265260'>ab265260</a>)

Lane 2:

FAS knockout HeLa cell lysate at 20 µg

Predicted band size: 37 kDa

Observed band size: 37 kDa

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• WB

Lab

Western blot - Anti-Fas antibody [EPR5700] (AB133619)

Lanes 1 - 4 : Merged signal (red and green). Green - ab133619 observed at 42 kDa. Red - loading control, ab8245, observed at 38 kDa.

This western blot image is a comparison between ab133619 and a competitor's top cited rabbit polyclonal antibody.

All lanes:

Western blot - Anti-Fas antibody [EPR5700] (ab133619)

Lane 1:

Wild-type HAP1 cell lysate at 20 µg

Lane 2:

Fas knockout HAP1 cell lysate at 20 µg

Lane 3:

Raji cell lysate at 20 µg

Lane 4:

Ramos cell lysate at 20 µg

Predicted band size: 37 kDa

false

• WB

Unknown

Western blot - Anti-Fas antibody [EPR5700] (AB133619)

Actual band size : 45 kDa

All lanes:

Western blot - Anti-Fas antibody [EPR5700] (ab133619) at 1/1000 dilution

Lane 1:

Ramos cell lysate at 10 µg

Lane 2:

HT-1080 cell lysate at 10 µg

Lane 3:

Raji cell lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 37 kDa

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