Anti-CD97 antibody [EPR4427]

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-CD97 antibody [EPR4427] (AB108368)

ICC/IF image of ab108368 stained MDA-MB-231 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab108368 at 1/200 dilution overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43μM.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD97 antibody [EPR4427] (AB108368)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling CD97 with ab108368 at 1/200 dilution.

Positive staining on human colon

The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument

Counterstained with Hematoxylin.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD97 antibody [EPR4427] (AB108368)

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling CD97 with ab108368 at 1/2000 dilution.

Positive staining on human spleen

The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument

Counterstained with Hematoxylin.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• Flow Cyt

Lab

Flow Cytometry - Anti-CD97 antibody [EPR4427] (AB108368)

Flow Cytometry analysis of MDA-MB-231 (human mammary gland epithelial adenocarcinoma) cells labeling CD97 with purified ab108368 at 1/250 dilution (10ug/ml) (red). A Goat anti rabbit IgG (Alexa Fluor^® 488)(1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

• IP

Lab

Immunoprecipitation - Anti-CD97 antibody [EPR4427] (AB108368)

CD97 was immunoprecipitated from 0.35 mg U-937 (Human histiocytic lymphoma monocyte) whole cell lysate 10 µg with ab108368 at 1/50 dilution (2µg). VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : U-937 (Human histiocytic lymphoma monocyte) whole cell lysate 10 µg

Lane 2 : ab108368 IP in U-937 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab108368 in U-937 whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-CD97 antibody [EPR4427] (ab108368)

Predicted band size: 92 kDa

Observed band size: 74-89 kDa

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• WB

Lab

Western blot - Anti-CD97 antibody [EPR4427] (AB108368)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/1000000 dilution.

Negative sample : HT-1080, HEK-293 (PMID : 30728423, PMID : 34028660)

All lanes:

Western blot - Anti-CD97 antibody [EPR4427] (ab108368) at 1/1000 dilution

Lane 1:

U-937 (human histiocytic lymphoma monocyte) whole cell lysate at 20 µg

Lane 2:

HT-1080 (human fibrosarcoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

HEK-293 (human embryonic kidney epithelial cell) whole cell lysate at 20 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

Observed band size: 92 kDa

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Exposure time: 40s

• WB

Unknown

Western blot - Anti-CD97 antibody [EPR4427] (AB108368)

All lanes:

Western blot - Anti-CD97 antibody [EPR4427] (ab108368) at 1/1000 dilution

Lane 1:

Human tonsil tissue lysate at 10 µg

Lane 2:

Jurkat cell lysate at 10 µg

Lane 3:

U937 (Human histiocytic lymphoma cell line) cell lysate at 10 µg

Lane 4:

Human fetal thymus tissue lysate at 10 µg

Predicted band size: 92 kDa

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