Anti-Beclin 1 antibody [EPR19662] - BSA and Azide free

• IP

Collaborator

Immunoprecipitation - Anti-Beclin 1 antibody [EPR19662] - BSA and Azide free (AB227107)

This data was developed using ab207612, the same antibody clone in a different buffer formulation.

Immunoprecipitation of BECN1 in HeLa cells. Lysates were prepared and immunoprecipitation was performed using 2 μg of ab207612 pre-coupled to Protein A beads. Samples were then washed and processed for western blot.

This data was 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.

false

• IP

Supplier Data

Immunoprecipitation - Anti-Beclin 1 antibody [EPR19662] - BSA and Azide free (AB227107)

Beclin 1 was immunoprecipitated from 0.35 mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab207612 at 1/30 dilution.

Western blot was performed from the immunoprecipitate using ab207612 at 1/1000 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1 : HeLa whole cell lysate 10μg (Input).

Lane 2 : ab207612 IP in HeLa whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab207612 in HeLa whole cell lysate.

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

Exposure time : 30 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab207612).

All lanes:

Immunoprecipitation - Anti-Beclin 1 antibody [EPR19662] (<a href='/en-us/products/primary-antibodies/beclin-1-antibody-epr19662-ab207612'>ab207612</a>)

Predicted band size: 52 kDa

false

• WB

Lab

Western blot - Anti-Beclin 1 antibody [EPR19662] - BSA and Azide free (AB227107)

Western blot : Anti-BECN1 antibody [EPR19662] (ab207612) staining at 1/2000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab207612 was shown to bind specifically to BECN1. A band was observed at 57 kDa in wild-type HeLa cell lysates with no signal observed at this size in BECN1 knockout cell line. To generate this image, wild-type and BECN1 knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-Beclin 1 antibody [EPR19662] (<a href='/en-us/products/primary-antibodies/beclin-1-antibody-epr19662-ab207612'>ab207612</a>) at 1/2000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

BECN1 knockout HeLa cell lysate at 20 µg

Lane 3:

Jurkat cell lysate at 20 µg

Lane 4:

HepG2 cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

false

• WB

Collaborator

Western blot - Anti-Beclin 1 antibody [EPR19662] - BSA and Azide free (AB227107)

This data was developed using ab207612, the same antibody clone in a different buffer formulation.

ab207612 was shown to react with BECN1 in wild-type HeLa cells in Western blot with loss of signal observed in BECN1 knockout cell line ab262511. Wild-type HeLa and BECN1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab207612 overnight at 4 °C at a 1/2000 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.

This data was 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-Beclin 1 antibody [EPR19662] (<a href='/en-us/products/primary-antibodies/beclin-1-antibody-epr19662-ab207612'>ab207612</a>) at 1/2000 dilution

Lane 1:

Wild-type HeLa lysate at 30 µg

Lane 2:

BECN1 knock-out HeLa lysate at 30 µg

Lane 2:

Western blot - Human BECN1 (Beclin 1) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-becn1-beclin-1-knockout-hela-cell-line-ab262511'>ab262511</a>) at 30 µg

false