Anti-IKK alpha antibody [EPR464] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IKK alpha antibody [EPR464] - BSA and Azide free (AB171362)

This data was developed using ab109749, the same antibody clone in a different buffer formulation.

ab109749, at 1/100 dilution, staining IKK alpha in paraffin-embedded Human breast tissue by Immunohistochemistry.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IP

Lab

Immunoprecipitation - Anti-IKK alpha antibody [EPR464] - BSA and Azide free (AB171362)

This data was developed using ab109749, the same antibody clone in a different buffer formulation.

IKK alpha was immunoprecipitated from 0.35 mg Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate 10 μg with 109749 at 1/50 dilution (2μg). VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate 10 μg

Lane 2 : ab109749 IP in Jurkat whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab109749 in Jurkat whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-IKK alpha antibody [EPR464] (<a href='/en-us/products/primary-antibodies/ikk-alpha-antibody-epr464-ab109749'>ab109749</a>)

Predicted band size: 84 kDa

Observed band size: 85 kDa

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• WB

Unknown

Western blot - Anti-IKK alpha antibody [EPR464] - BSA and Azide free (AB171362)

This data was developed using ab109749, the same antibody clone in a different buffer formulation.

Lanes 1, 3, and 5 : ild-type HAP1 cell lysate (20 μg)

Lanes 2, 4, and 6 : IKK alpha knockout HAP1 cell lysate (20 μg)

Lanes 1 and 2 : Green signal from target - ab109749 observed at 84 kDa

Lanes 3 and 4 : Red signal from loading control - ab8226 observed at 42 kDa

Lanes 5 and 6 : Merged (red and green) signal

ab109749 was shown to specifically react with IKK alpha when IKK alpha knockout samples were used. Wild-type and IKK alpha knockout samples were subjected to SDS-PAGE. ab109749 and ab8226 (loading control to beta actin) were both diluted at 1/1000 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 1/10000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-IKK alpha antibody [EPR464] (<a href='/en-us/products/primary-antibodies/ikk-alpha-antibody-epr464-ab109749'>ab109749</a>)

Predicted band size: 84 kDa

false

• WB

Unknown

Western blot - Anti-IKK alpha antibody [EPR464] - BSA and Azide free (AB171362)

This data was developed using ab109749, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-IKK alpha antibody [EPR464] (<a href='/en-us/products/primary-antibodies/ikk-alpha-antibody-epr464-ab109749'>ab109749</a>) at 1/1000 dilution

Lane 1:

Jurkat cell lysate at 10 µg

Lane 2:

HeLa cell lysate at 10 µg

Lane 3:

HepG2 cell lysate at 10 µg

Lane 4:

Daudi cell lysate at 10 µg

Predicted band size: 84 kDa

false