Anti-MVP antibody [EPR23594-106]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-MVP antibody [EPR23594-106] (AB273093)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized A549 (Human lung carcinoma epithelial cell) cells labelling MVP with ab273093 at 1/50 dilution (1μg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MVP antibody [EPR23594-106] (AB273093)

Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling MVP with ab273093 at 1/4000 dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in endothelium of human cerebrum (PMID : 14636345). Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-MVP antibody [EPR23594-106] (AB273093)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% Triton X-100 permeabilized A549 cells labelling MVP with ab273093 at 1/50 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 2 μg/ml dilution (Green). Confocal image showing cytoplasmic staining in A549 cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 2.5 μg/ml dilution (Red). The nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 2 μg/ml dilution.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MVP antibody [EPR23594-106] (AB273093)

Immunohistochemical analysis of paraffin-embedded human ovary cancer tissue labeling MVP with ab273093 at 1/4000 dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in human ovary cancer (PMID : 23739867). Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MVP antibody [EPR23594-106] (AB273093)

Immunohistochemical analysis of paraffin-embedded human lung cancer tissue labeling MVP with ab273093 at 1/4000 dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in human lung cancer (PMID : 22117969). Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-MVP antibody [EPR23594-106] (AB273093)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 cells labelling MVP with ab273093 at 1/50 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 2 μg/ml dilution (Green). Confocal image showing cytoplasmic staining in NIH/3T3 cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 2.5 μg/ml dilution (Red). The nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 2 μg/ml dilution.

• WB

Lab

Western blot - Anti-MVP antibody [EPR23594-106] (AB273093)

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

The molecular weight observed is consistent with what has been described in the literature (PMID : 23462207).

MVP expression can be induced by IFN gamma in HeLa cell (PMID : 16418217).

Exposure time : 70 seconds.

All lanes:

Western blot - Anti-MVP antibody [EPR23594-106] (ab273093) at 1/1000 dilution

Lane 1:

A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

A431 (human epidermoid carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

Caco-2 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

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

Lane 5:

HeLa treated with 100U/ml IFN gamma for 24 hours, whole cell lysate at 20 µg

Lane 6:

RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 7:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 99 kDa

Observed band size: 110 kDa

false

• WB

Lab

Western blot - Anti-MVP antibody [EPR23594-106] (AB273093)

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

Exposure time : 10 seconds.

All lanes:

Western blot - Anti-MVP antibody [EPR23594-106] (ab273093) at 1/1000 dilution

Lane 1:

Human colon tissue lysate at 20 µg

Lane 2:

Human lung tissue lysate at 20 µg

Lane 3:

Human kidney tissue lysate at 20 µg

Secondary

All lanes:

VeriBlot for IP secondary antibody(HRP)(<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Predicted band size: 99 kDa

Observed band size: 110 kDa

false

• WB

Lab

Western blot - Anti-MVP antibody [EPR23594-106] (AB273093)

Lanes 1-3 : Merged signal (red and green). Green - ab273093 observed at 110 kDa. Red - loading control ab8245 (Mouse monoclonal [6C5] to GAPDH) observed at 36 kDa.

ab273093 Anti-MVP antibody [EPR23594-106] was shown to specifically react with MVP in wild-type HeLa cells in Western blot. Significant decrease (8.7 % of intensity compared to the WT band) of signal was observed when MVP knockout cell line ab264817 (knockout cell lysate ab257544) was used.

ab273093 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at 4℃ overnight 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-MVP antibody [EPR23594-106] (ab273093) at 1/1000 dilution

Lane 1:

Wild-type HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

MVP knockout HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

Western blot - Human MVP knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-mvp-knockout-hela-cell-line-ab264817'>ab264817</a>)

Lane 3:

Caco-2 (human colorectal adenocarcinoma epithelial cell) whole 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: 99 kDa

Observed band size: 110 kDa

false

• WB

Lab

Western blot - Anti-MVP antibody [EPR23594-106] (AB273093)

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

Exposure time : Lane 1 : 26 seconds; Lane 2-4 : 70 seconds.

All lanes:

Western blot - Anti-MVP antibody [EPR23594-106] (ab273093) at 1/1000 dilution

Lane 1:

Mouse colon tissue lysate at 20 µg

Lane 2:

Mouse kidney tissue lysate at 20 µg

Lane 3:

C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

Lane 4:

PC-12 (rat adrenal gland pheochromocytoma ) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 99 kDa

Observed band size: 110 kDa

false