Anti-IL-6 antibody [EPR21710]

• IP

Supplier Data

Immunoprecipitation - Anti-IL-6 antibody [EPR21710] (AB229381)

IL6 was immunoprecipitated from 0.35 mg of RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) treated with 100 ng/ml Lipopolysaccharide (LPS) for 6 hours, then added 300 ng/ml BFA the last 3 hours, whole cell lysate with ab229381 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab229381 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

Lane 1 : RAW 264.7 treated with 100 ng/ml Lipopolysaccharide (LPS) for 6 hours, then added 300 ng/ml BFA the last 3 hours, whole cell lysate 10 μg (Input).
Lane 2 : ab229381 IP in RAW 264.7 treated with 100 ng/ml Lipopolysaccharide (LPS) for 6 hours, then added 300 ng/ml BFA the last 3 hours, whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab229381 in RAW 264.7 treated with 100 ng/ml Lipopolysaccharide (LPS) for 6 hours, then added 300 ng/ml BFA the last 3 hours, whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 2 minutes.

All lanes:

Immunoprecipitation - Anti-IL-6 antibody [EPR21710] (ab229381)

Predicted band size: 23 kDa

Observed band size: 24 kDa

true

• WB

Supplier Data

Western blot - Anti-IL-6 antibody [EPR21710] (AB229381)

Western blot analysis with ab229381 (left) and control product ab259341 (right).

Blocking and diluting buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-IL-6 antibody [EPR21710] (ab229381) at 1/1000 dilution

Lanes 1 and 3:

Untreated RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lanes 2 and 4:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) treated with 100ng/ml lipopolysaccharide (LPS) for 6 hours, then added 300 ng/ml BFA for the last 3 hours, whole cell lysate at 20 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 1/20000 dilution

Predicted band size: 23 kDa

Observed band size: 24 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-IL-6 antibody [EPR21710] (AB229381)

Blocking/Dilution buffer : 5% NFDM/TBST.

We detect a non-specific band at approximately 60 KDa.

All lanes:

Western blot - Anti-IL-6 antibody [EPR21710] (ab229381) at 1/1000 dilution

Lane 1:

Untreated RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus), whole cell lysate at 10 µg

Lane 2:

RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) treated with 100 ng/ml Lipopolysaccharides (LPS) for 6 hours, then added 300 ng/ml BFA for the last 3 hours, whole cell lysate, 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: 23 kDa

Observed band size: 24 kDa

true

Exposure time: 3min