Anti-c-Fos antibody [EPR20769]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-c-Fos antibody [EPR20769] (AB214672)

Immunofluorescent analysis of 4% paraformaldehyde-fixed. 0.1% Triton X-100 permeabilized serum treated and non-treated HeLa (human cervix adenocarcinoma epithelial cell) cells labeling c-Fos with ab214672 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing weakly nuclear staining on HeLa cells grown in serum free medium for 36 hours. Expression of c-Fos increased in HeLa cells grown in serum free medium for 36 hours followed by addition of 20% FBS for 2 hours.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) (red) at 1/200 dilution.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-c-Fos antibody [EPR20769] (AB214672)

c-Fos was immunoprecipitated from 0.35 mg of HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab214672 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab214672 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution
Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) grown in serum free medium for 36 hours, followed by addition of 20%FBS for 2 hours, whole cell lysate, 10 μg (Input).
Lane 2 : ab214672 IP in HeLa grown in serum free medium for 36 hours, followed by addition of 20% FBS for 2 hours, whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab214672 in HeLa grown in serum free medium for 36 hours, followed by addition of 20% FBS for 2 hours, whole cell lysate.
Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 1 second.

The observed lower band is a proteasomal degradation fragment (PMID : 9737957).

All lanes:

Immunoprecipitation - Anti-c-Fos antibody [EPR20769] (ab214672)

Predicted band size: 41 kDa

Observed band size: 55-60 kDa

true

• WB

Supplier Data

Western blot - Anti-c-Fos antibody [EPR20769] (AB214672)

Exposure time : 15 seconds.

Blocking/Dilution buffer : 5% NFDM/TBST.

The expression of c-Fos is induced by the addition of 20%FBS (PMID : 24386331, PMID : 23300800, PMID : 25695333).

All lanes:

Western blot - Anti-c-Fos antibody [EPR20769] (ab214672) at 1/1000 dilution

Lane 1:

Untreated HeLa (human epithelial cell line from cervix adenocarcinoma) grown in serum free medium for 36 hours, whole cell lysate at 10 µg

Lane 2:

HeLa (human epithelial cell line from cervix adenocarcinoma) grown in serum free medium for 36 hours, followed by addition of 20% FBS for 2 hours, whole cell lysate at 10 µg

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 1/50000 dilution

Predicted band size: 41 kDa

Observed band size: 55-60 kDa

true

• WB

Supplier Data

Western blot - Anti-c-Fos antibody [EPR20769] (AB214672)

Exposure time : 3 minutes.

Blocking/Dilution buffer : 5% NFDM/TBST.

PMA treatment induces expression of c-Fos, as documented in the literature (PMID : 24386331, PMID : 23300800, PMID : 25695333).

All lanes:

Western blot - Anti-c-Fos antibody [EPR20769] (ab214672) at 1/5000 dilution

Lane 1:

Untreated Jurkat (human T cell leukemia cell line from peripheral blood) grown in serum free medium overnight, whole cell lysate at 10 µg

Lane 2:

Jurkat (human T cell leukemia cell line from peripheral blood) grown in serum free medium overnight, followed by treatment with 200 nM PMA for 4 hours, whole cell lysate at 10 µg

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 1/100000 dilution

Predicted band size: 41 kDa

Observed band size: 55-60 kDa

true

• WB

Supplier Data

Western blot - Anti-c-Fos antibody [EPR20769] (AB214672)

Exposure time : 1 minute.

Blocking/Dilution buffer : 5% NFDM/TBST.

PMA treatment induces expression of c-Fos, as documented in the literature (PMID : 24386331, PMID : 23300800, PMID : 25695333).

All lanes:

Western blot - Anti-c-Fos antibody [EPR20769] (ab214672) at 1/5000 dilution

Lane 1:

Untreated RAW264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus grown in serum free medium overnight, whole cell lysate at 10 µg

Lane 2:

RAW264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) grown in serum free medium overnight, followed by treatment with 200 nM PMA for 4 hours, whole cell lysate at 10 µg

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 1/100000 dilution

Predicted band size: 41 kDa

Observed band size: 55-60 kDa

true