Anti-IL-17F antibody [EPR17830-169]

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-IL-17F antibody [EPR17830-169] (AB187059)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed 90% methanol-fixed EL4 (mouse lymphoma T lymphocyte) treated with 50ng/ml Phorbol-12-myristate-13-acetate (PMA) and 500ng/ml Ionomycin calcium salt for 24 hours, and 500ng/ml Brefeldin A (BFA) was added for the last 20 hours (Right) / Untreated control (Left) labeling IL17F with ab187059 at 1/60 dilution.

Secondary antibody used Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at a 1/2000 dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-IL-17F antibody [EPR17830-169] (AB187059)

IL17F was immunoprecipitated from 0.35 mg of EL4 (mouse lymphoma T lymphocyte) treated with 50 ng/ml phorbol-12-myristate-13-acetate (PMA) and 500 ng/ml ionomycin calcium salt for 24 hours, and 500ng/ml Brefeldin A (BFA) was added for the last 20 hours whole cell lysate with ab187059 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab187059 at 1/1,000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/5,000 dilution.

Lane 1 : EL4 (mouse lymphoma T lymphocyte) treated with 50 ng/ml phorbol-12-myristate-13-acetate (PMA) and 500 ng/ml ionomycin calcium salt for 24 hours, and 500ng/ml Brefeldin A (BFA) was added for the last 20 hours whole cell lysate 10 μg (Input).
Lane 2 : EL4 treated with 50 ng/ml phorbol-12-myristate-13-acetate (PMA) and 500 ng/ml ionomycin calcium salt for 24 hours, and 500 ng/ml Brefeldin A (BFA) was added for the last 20 hours whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab187059 in EL4 treated with 50 ng/ml phorbol-12-myristate-13-acetate (PMA) and 500 ng/ml ionomycin calcium salt for 24 hours, and 500 ng/ml Brefeldin A (BFA) was added for the last 20 hours whole cell lysate.

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : 10 seconds

All lanes:

Immunoprecipitation - Anti-IL-17F antibody [EPR17830-169] (ab187059)

Predicted band size: 18 kDa

false

• WB

Supplier Data

Western blot - Anti-IL-17F antibody [EPR17830-169] (AB187059)

Blocking/Diluting buffer : 5% NFDM/TBST

The molecular mass observed is consistent with the literature (PMID 2212322).

Expression of IL-17F in EL4 cells is increased by PMA and Ionomycin treatment (PMID 28382171).

All lanes:

Western blot - Anti-IL-17F antibody [EPR17830-169] (ab187059) at 1/1000 dilution

Lane 1:

Untreated EL4 (mouse lymphoma T lymphocyte), whole cell lysate at 10 µg

Lane 2:

EL4 treated with 50 ng/ml phorbol-12-myristate-13-acetate (PMA) and 500 ng/ml ionomycin calcium salt for 24 hours, and 500 ng/ml Brefeldin A (BFA) was added to the treated cells last 20 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: 18 kDa

Observed band size: 16-18 kDa

false

Exposure time: 48s

• WB

Supplier Data

Western blot - Anti-IL-17F antibody [EPR17830-169] (AB187059)

Blocking/Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-IL-17F antibody [EPR17830-169] (ab187059) at 1/5000 dilution

Lane 1:

Mouse IL17A recombinant protein (aa26-158) 10 ng

Lane 2:

Mouse IL17F recombinant protein (aa29-161) 10 ng

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: 18 kDa

Observed band size: 18 kDa

false

Exposure time: 26s