Anti-DDB2 antibody [EPR9811] - BSA and Azide free

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-DDB2 antibody [EPR9811] - BSA and Azide free (AB250373)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181136).

Immunohistochemical analysis of paraffin-embedded Rat skeletal muscle tissue labeling DCAMKL1 with ab202755 at 1/100 dilution.

Negative control : no staining on rat skeletal muscle.

The primary antibody was incubated for 60 mins at room temperature (shown in magenta). Nuclear DNA was labeled with DAPI (shown in blue). The section was then mounted using Fluoromount^®.

The immunostaining was performed on a Leica Biosystems BOND RX instrument.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Counterstained with DAPI.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 40 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DDB2 antibody [EPR9811] - BSA and Azide free (AB250373)

This data was developed using ab181136, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human stomach carcinoma tissue sections labeling DDB2 with Purified ab181136 at 1/2500 dilution (0.25 µg/mL). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• WB

Unknown

Western blot - Anti-DDB2 antibody [EPR9811] - BSA and Azide free (AB250373)

This data was developed using the same antibody clone in a different buffer formulation (ab181136).

40kDa band might be an isomer of DNA damage-binding protein 2 (Isomer D3).

All lanes:

Western blot - Anti-DDB2 antibody [EPR9811] (<a href='/en-us/products/primary-antibodies/ddb2-antibody-epr9811-ab181136'>ab181136</a>) at 1/2000 dilution

Lane 1:

SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate at 15 µg

Lane 2:

Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysate at 15 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: 48 kDa

false

• WB

Supplier Data

Western blot - Anti-DDB2 antibody [EPR9811] - BSA and Azide free (AB250373)

This data was developed using the same antibody clone in a different buffer formulation (ab181136).

40kDa band might be an isomer of DNA damage-binding protein 2 (Isomer D3).

All lanes:

Western blot - Anti-DDB2 antibody [EPR9811] (<a href='/en-us/products/primary-antibodies/ddb2-antibody-epr9811-ab181136'>ab181136</a>) at 1/1000 dilution

All lanes:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 15 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: 48 kDa

false

• WB

Lab

Western blot - Anti-DDB2 antibody [EPR9811] - BSA and Azide free (AB250373)

This data was developed using the same antibody clone in a different buffer formulation (ab181136).

Lanes 1-3 : Merged signal (red and green). Green - ab181136 observed at 48 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab181136 Anti-DDB2 antibody [EPR9811] was shown to specifically react with DDB2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265115 (knockout cell lysate ab257177) was used. Wild-type and DDB2 knockout samples were subjected to SDS-PAGE. ab181136 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. 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 in 10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-DDB2 antibody [EPR9811] (<a href='/en-us/products/primary-antibodies/ddb2-antibody-epr9811-ab181136'>ab181136</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

DDB2 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human DDB2 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-ddb2-knockout-hela-cell-line-ab265115'>ab265115</a>)

Lane 3:

Raji cell lysate at 20 µg

Predicted band size: 48 kDa

Observed band size: 48 kDa

false