Anti-DDB2 antibody [EPR9811]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DDB2 antibody [EPR9811] (AB181136)

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] (AB181136)

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

All lanes:

Western blot - Anti-DDB2 antibody [EPR9811] (ab181136) 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

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• WB

Supplier Data

Western blot - Anti-DDB2 antibody [EPR9811] (AB181136)

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

All lanes:

Western blot - Anti-DDB2 antibody [EPR9811] (ab181136) 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

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• WB

Unknown

Western blot - Anti-DDB2 antibody [EPR9811] (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] (ab181136) 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 3:

Raji cell lysate at 20 µg

Predicted band size: 48 kDa

Observed band size: 48 kDa

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• WB

Lab

Western blot - Anti-DDB2 antibody [EPR9811] (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] (ab181136) 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

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• WB

CiteAb

Western blot - Anti-DDB2 antibody [EPR9811] (AB181136)

DDB2 western blot using anti-DDB2 antibody [EPR9811] ab181136. Publication image and figure legend from Kumar, N., Theil, A. F., et al., 2022, Nat Commun, PubMed 35190564.

ab181136 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab181136 please see the product overview.

DDB2 binds sparse telomeric 8-oxoG independently of the DDB1-Cul4A-RBX1 E3 ligase.a Representative images showing recruitment of DDB2-mCherry to telomeric 8-oxoG in cells transfected with control, DDB1 or Cul4A siRNA. b Quantification of a. c, e DDB2-mCherry and GFP-DDB1 (c) or DDB2-mCherry and GFP-Cul4A (e) accumulation at 8-oxoG sites after dye (100 nM, 15 min) plus light (660 nm, 10 min) treatment. d, f Quantification of c and e respectively. g Western blot for DDB2 in U2OS-FAP-TRF1 cells treated with UVC, potassium bromate (KBrO3) or dye plus light at indicated doses. Independent experiments are represented by black circles. h Colocalization of DDB2-mCherry and GFP-Cul4A at damaged telomeres in U2OS-FAP-TRF1 cells transfected with control or OGG1 siRNA. i Quantification of h. Data (a–h) represents mean ± SEM from two independent experiments. 'n' represents the number of cells scored for each condition. One-way ANOVA (Sidak multiple comparison test) (b, i) was performed for statistical analysis : *p < 0.05, **p < 0.01, ***p < 0.001, ns Not significant. Scale : 5 μm. Source data are provided as a Source Data file. (See also Supplementary Fig. 5).

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