Anti-CDA antibody [EPR20525]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDA antibody [EPR20525] (AB222515)

Immunohistochemical analysis of paraffin-embedded human normal colon (panel A) and colon cancer (panel B) tissues labeling CDA with ab222515 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic and nuclear staining on sporadic stromal cells of human normal colon tissue, while adjacent cancer cells and some stromal cells show moderate positive staining. The IHC signal on human colon cancer tissue was higher than its corresponding normal tissue (PMID : 9849491). Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDA antibody [EPR20525] (AB222515)

Immunohistochemical analysis of paraffin-embedded human normal liver (panel A) and liver cancer (panel B) tissues labeling CDA with ab222515 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic and nuclear staining on normal human liver tissues, with only sporadic stromal cells showing positive staining in human liver cancer. The IHC signal on human liver cancer tissue was much lower than its corresponding normal tissue (PMID : 9849491). Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-CDA antibody [EPR20525] (AB222515)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HeLa (humanepithelialcell line from cervix adenocarcinoma) cell line labeling CDA with ab222515 at 1/70 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDA antibody [EPR20525] (AB222515)

Immunohistochemical analysis of paraffin-embedded human spleen tissue labeling CDA with ab222515 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear and cytoplasmic staining on neutrophils of human spleen (PMID : 11069255). Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-CDA antibody [EPR20525] (AB222515)

Western blot : Rabbit Monoclonal [EPR20525] to CDA ab222515 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH (ab8245) loading control staining at 1/20,000 dilution, shown in magenta.

A band was observed at 16 kDa in Wild-type A549 cell lysates with no signal observed at this size in CDA knockout A549 cell line.

To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc 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 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-CDA antibody [EPR20525] (ab222515) at 1/1000 dilution

Lane 1:

Wild-type A549 at 20 µg

Lane 2:

CDA knockout A549 at 20 µg

Lane 2:

Western blot - Human CDA knockout A549 cell line (<a href='/en-us/products/cell-lines/human-cda-knockout-a549-cell-line-ab300883'>ab300883</a>) at 20 µg

Lane 3:

Wild-type HCT at 20 µg

Lane 4:

CDA knockout HCT at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 16 kDa

Observed band size: 16 kDa

false

• WB

Lab

Western blot - Anti-CDA antibody [EPR20525] (AB222515)

Western blot : Rabbit Monoclonal [EPR20525] to CDA ab222515 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH (ab8245) loading control staining at 1/20,000 dilution, shown in magenta.

A band was observed at 17 kDa in Wild-type HCT 116 cell lysates with no signal observed at this size in cda knockout HCT 116 cell line.

To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc 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 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-CDA antibody [EPR20525] (ab222515) at 1/1000 dilution

Lane 1:

Wild-type HCT 116 at 20 µg

Lane 2:

CDA knockout HCT 116 at 20 µg

Lane 2:

Western blot - Human CDA knockout HCT116 cell line (ab300882) at 20 µg

Lane 3:

HepG2 at 20 µg

Lane 4:

MCF7 at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 16 kDa

Observed band size: 17 kDa

false

• WB

Supplier Data

Western blot - Anti-CDA antibody [EPR20525] (AB222515)

Exposure times : Lane 1 : 3 minutes; Lane 2 : 8 seconds; Lanes 3/5 : 5 seconds; Lane 4 : 30 seconds.

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-CDA antibody [EPR20525] (ab222515) at 1/1000 dilution

Lane 1:

HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 10 µg

Lane 2:

Human fetal spleen lysate at 10 µg

Lane 3:

HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate at 10 µg

Lane 4:

TF-1 (human bone marrow erythroleukemia cell line) whole cell lysate at 10 µg

Lane 5:

Human fetal kidney lysate at 10 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: 16 kDa

Observed band size: 16 kDa

true