Anti-CD147 antibody [EPR4053] - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD147 antibody [EPR4053] - BSA and Azide free (AB247626)

This data was developed using ab108308, the same antibody clone in a different buffer formulation.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon cancer tissue sections labeling CD147 with purified ab108308 at 1/3000 dilution (0.1 μg/mL). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• Flow Cyt

Lab

Flow Cytometry - Anti-CD147 antibody [EPR4053] - BSA and Azide free (AB247626)

This data was developed using ab108308, the same antibody clone in a different buffer formulation.
Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling CD147 with purified ab108308 at 1/30 dilution (10 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CD147 antibody [EPR4053] - BSA and Azide free (AB247626)

This data was developed using ab108308, the same antibody clone in a different buffer formulation.

Immunocytochemistry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling CD147 with purified ab108308 at 1/50 dilution (6.3 μg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1/200 (2.5 μg/mL). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/1000 (2 μg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• WB

Lab

Western blot - Anti-CD147 antibody [EPR4053] - BSA and Azide free (AB247626)

Lanes 1-4 : Merged signal (red and green). Green - ab108308 observed at 50 kDa. Red - loading control ab8245 observed at 36 kDa.

ab108308 Anti-CD147 antibody [EPR4053] was shown to specifically react with CD147 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266331 (knockout cell lysate ab256853) was used. Wild-type and CD147 knockout samples were subjected to SDS-PAGE. ab108308 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 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-CD147 antibody [EPR4053] (<a href='/en-us/products/primary-antibodies/cd147-antibody-epr4053-ab108308'>ab108308</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

BSG knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human BSG (CD147) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-bsg-cd147-knockout-hek-293t-cell-line-ab266331'>ab266331</a>)

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

U-87 MG cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 42 kDa,65 kDa

Observed band size: 50 kDa,65 kDa

false

• WB

Lab

Western blot - Anti-CD147 antibody [EPR4053] - BSA and Azide free (AB247626)

This data was developed using the same antibody clone in a different buffer formulation (ab108308).

Lanes 1 - 3 : Merged signal (red and green). Green - ab108308 observed at 42-70 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.

ab108308 was shown to react with CD147 in wild-type A549 cells in western blot with loss of signal observed in BSG knockout cell line ab273748 (knockout cell lysate ab275500). Wild-type and BSG knockout A549 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab108308 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-CD147 antibody [EPR4053] (<a href='/en-us/products/primary-antibodies/cd147-antibody-epr4053-ab108308'>ab108308</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 30 µg

Lane 2:

BSG knockout A549 cell lysate at 30 µg

Lane 2:

Western blot - Human BSG (CD147) knockout A549 cell line (<a href='/en-us/products/cell-lines/human-bsg-cd147-knockout-a549-cell-line-ab273748'>ab273748</a>)

Lane 3:

Raji cell lysate at 30 µg

Predicted band size: 42 kDa

Observed band size: 42-70 kDa

false

• WB

Unknown

Western blot - Anti-CD147 antibody [EPR4053] - BSA and Azide free (AB247626)

All lanes:

Western blot - Anti-CD147 antibody [EPR4053] (<a href='/en-us/products/primary-antibodies/cd147-antibody-epr4053-ab108308'>ab108308</a>) at 1/1000 dilution

Lane 1:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

U-87 MG (Human glioblastoma-astrocytoma 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

Predicted band size: 42 kDa

false