Anti-NAK/TBK1 antibody [EPR2867(2)-19] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NAK/TBK1 antibody [EPR2867(2)-19] - BSA and Azide free (AB186468)

ab109735 staining NAK/TBK1 in Human liver tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed and paraffin-embedded, antigen retrieval was by heat mediation in Tris/EDTA buffer pH9. Samples were incubated with primary antibody (1/800). An undiluted HRP-conjugated anti-rabbit IgG was used as the secondary antibody. Tissue counterstained with Hematoxylin.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109735).

• IP

Unknown

Immunoprecipitation - Anti-NAK/TBK1 antibody [EPR2867(2)-19] - BSA and Azide free (AB186468)

This data was developed using ab109735, the same antibody clone in a different buffer formulation :

Immunoprecipitation of TBK1 in HeLa cells. Lysates were prepared and immunoprecipitation was performed using 1.0 μg of ab109735 pre-coupled to prot.A-Sepharose beads. Samples were washed and processed for western blot with ab12116 at 1 in 2000. This data was kindly provided by the YCharOS Inc., an open science company with the mission of characterizing every commercially available antibody reagent. Abcam are working with YCharOS to support their mission of antibody characterisation using knock out cell lines.

All lanes:

Immunoprecipitation - Anti-NAK/TBK1 antibody [EPR2867(2)-19] - BSA and Azide free (ab186468)

Predicted band size: 84 kDa

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• IP

Unknown

Immunoprecipitation - Anti-NAK/TBK1 antibody [EPR2867(2)-19] - BSA and Azide free (AB186468)

Purified ab109735 at 1/30 dilution (2μg) immunoprecipitating NAK/TBK1 in HeLa whole cell lysate.
Lane 1 (input) : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10μg
Lane 2 (+) : ab109735 + HeLa whole cell lysate.
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab109735 in HeLa whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/5000 dilution) was used for Western blotting.
Blocking Buffer and concentration : 5% NFDM/TBST.
Diluting buffer and concentration : 5% NFDM/TBST.
Observed band size : 84 kDa
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109735).

All lanes:

Immunoprecipitation - Anti-NAK/TBK1 antibody [EPR2867(2)-19] (<a href='/en-us/products/primary-antibodies/nak-tbk1-antibody-epr28672-19-ab109735'>ab109735</a>)

Predicted band size: 61 kDa,84 kDa,94 kDa

Observed band size: 85 kDa

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• WB

Unknown

Western blot - Anti-NAK/TBK1 antibody [EPR2867(2)-19] - BSA and Azide free (AB186468)

This data was developed using ab109735, the same antibody clone in a different buffer formulation

All lanes:

Western blot - Anti-NAK/TBK1 antibody [EPR2867(2)-19] - BSA and Azide free (ab186468) at 1/1000 dilution

Lane 1:

HeLa cell lysate at 10 µg

Lane 2:

U937 cell lysate at 10 µg

Lane 3:

HepG2 cell lysate at 10 µg

Lane 4:

293T cell lysate at 10 µg

Predicted band size: 84 kDa

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• WB

Lab

Western blot - Anti-NAK/TBK1 antibody [EPR2867(2)-19] - BSA and Azide free (AB186468)

This data was developed using ab109735, the same antibody clone in a different buffer formulation :

Lane 1 : Wild-type HAP1 cell lysate (20 μg)
Lane 2 : NAK/TBK1 knockout HAP1 cell lysate (20 μg)
Lane 3 : HeLa cell lysate (20 μg)
Lane 4 : HepG2 cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab109735 observed at 90 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab109735 was shown to recognize NAK/TBK1 when NAK/TBK1 knockout samples were used, along with additional cross-reactive bands. Wild-type and NAK/TBK1 knockout samples were subjected to SDS-PAGE. ab109735 and ab8245 (loading control to GAPDH) were diluted at 1/1000 and 1/10 000 respectively 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-NAK/TBK1 antibody [EPR2867(2)-19] - BSA and Azide free (ab186468)

Predicted band size: 84 kDa

false

• WB

Unknown

Western blot - Anti-NAK/TBK1 antibody [EPR2867(2)-19] - BSA and Azide free (AB186468)

This data was developed using ab109735, the same antibody clone in a different buffer formulation :

ab109735 was shown to react with TBK1 in wild-type U2OSn cells in Western blot with loss of signal observed in a TBK1 knockout cell line. Wild-type U2OSn and TBK1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab109735 overnight at 4 °C at a 1/10000 dilution. Blots were incubated with goat anti-rabbit HRP secondary antibodies at 1/5000 before imaging. These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

All lanes:

Western blot - Anti-NAK/TBK1 antibody [EPR2867(2)-19] - BSA and Azide free (ab186468)

Predicted band size: 84 kDa

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