Anti-CD147 antibody [EPR18008-67]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-CD147 antibody [EPR18008-67] (AB212057)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% TritonX-100 permeabilized bEnd.3 (mouse brain endothelioma cell line) cells labeling CD147 with ab212057 at 1/250 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cell membranous staining on bEnd.3 cell line.

The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/1000 dilution (red).

The negative control are is as follows :

-ve control 1 : ab212057 at 1/250 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/1000 dilution.
-ve control 2 : ab7291 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.

• Flow Cyt

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Flow Cytometry - Anti-CD147 antibody [EPR18008-67] (AB212057)

Flow cytometric analysis of fresh mouse thymocytes labeling CD147 with ab212057 at 1/200 dilution (red) compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (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 1/500 dilution was used as the secondary antibody.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-CD147 antibody [EPR18008-67] (AB212057)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% TritonX-100 permeabilized WEHI-231 (mouse lymphoblast B cell lymphoma cell line) cells labeling CD147 with ab212057 at 1/250 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cell membranous staining on bEnd.3 cell line.

The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution (red).

The negative control are is as follows :

-ve control 1 : ab212057 at 1/250 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution.
-ve control 2 : ab7291 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD147 antibody [EPR18008-67] (AB212057)

Immunohistochemical analysis of paraffin-embedded mouse intestine tissue labeling CD147 with ab212057 at 1/8000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Membranous staining on mouse intestine is observed. Counter stained withematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

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

• IP

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Immunoprecipitation - Anti-CD147 antibody [EPR18008-67] (AB212057)

CD147 was immunoprecipitated from 1 mg of RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate with ab212057 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab212057 at 1/5000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution

Lane 1 : RAW 264.7 whole cell lysate 10μg (Input).

Lane 2 : ab212057 IP in RAW 264.7 whole cell lysate.

Lane 3 : : Rabbit monoclonal IgG (ab172730) instead of ab212057 in RAW 264.7 whole cell lysate.

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

Exposure time : 1 second.

All lanes:

Immunoprecipitation - Anti-CD147 antibody [EPR18008-67] (ab212057)

Predicted band size: 42 kDa

Observed band size: 55 kDa

true

• WB

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Western blot - Anti-CD147 antibody [EPR18008-67] (AB212057)

Exposure time : Lanes 1/2 : 3 minutes; Lane 3 : 10 seconds; Lane 4 : 1 second.

Blocking and dilution buffer : 5% NFDM/TBST.

The expression profile observed is consistent with what has been described in the literature (PMID : 16721788; PMID : 23966157).

All lanes:

Western blot - Anti-CD147 antibody [EPR18008-67] (ab212057) at 1/5000 dilution

Lane 1:

RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg

Lane 2:

Mouse liver lysate at 10 µg

Lane 3:

WEHI-3 (mouse leukemia cell line) whole cell lysate at 10 µg

Lane 4:

bEnd.3 (mouse brain endothelioma cell line) whole cell 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 1/100000 dilution

Predicted band size: 42 kDa

Observed band size: 45-55 kDa

true

• WB

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Western blot - Anti-CD147 antibody [EPR18008-67] (AB212057)

Blocking and dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-CD147 antibody [EPR18008-67] (ab212057) at 1/5000 dilution

All lanes:

His-tagged mouse CD147 recombinant protein fragment (aa140-325) at 0.002 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 1/100000 dilution

Predicted band size: 42 kDa

Observed band size: 28 kDa

true

Exposure time: 3min