Anti-MVP antibody [EPR13227(B)]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-MVP antibody [EPR13227(B)] (AB175239)

Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling MVP with purified ab175239 at a dilution of 1/150. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

Control 1 : primary antibody (1/150) and secondary antibody, ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000).

Control 2 : ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MVP antibody [EPR13227(B)] (AB175239)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human pancreas tissue labeling MVP with unpurified ab175239 at a dilution of 1/50.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-MVP antibody [EPR13227(B)] (AB175239)

Immunocytochemistry/Immunofluorescence analysis of A549 cells labeling MVP with unpurified ab175239 at a dilution of 1/250 (red) and DAPI staining (blue).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MVP antibody [EPR13227(B)] (AB175239)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue labeling MVP with unpurified ab175239 at a dilution of 1/50.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-MVP antibody [EPR13227(B)] (AB175239)

Intracellular Flow Cytometry analysis of A549 cells labelling MVP with purified ab175239 at a dilution of 1/180 (red). Cells were fixed with 4% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-MVP antibody [EPR13227(B)] (AB175239)

Intracellular flow cytometrical analysis of permeabilized A549 cells labeling MVP with unpurified ab175239 antibody at a dilution of 1/100 (red) compared to a negative control (Rabbit IgG, green).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MVP antibody [EPR13227(B)] (AB175239)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue labelling MVP with purified ab175239 at a dilution of 1/350. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

• WB

Supplier Data

Western blot - Anti-MVP antibody [EPR13227(B)] (AB175239)

All lanes:

Western blot - Anti-MVP antibody [EPR13227(B)] (ab175239) at 1/10000 dilution

Lane 1:

A549 cell lysate at 10 µg

Lane 2:

Calu-3 cell lysate at 10 µg

Lane 3:

HeLa cell lysate at 10 µg

Predicted band size: 99 kDa

Observed band size: 110 kDa

false

• WB

Lab

Western blot - Anti-MVP antibody [EPR13227(B)] (AB175239)

ab175239 Anti-MVP antibody [EPR13227(B)] was shown to specifically react with MVP in HeLa wild-type cells ( ab255448). Loss of signal was observed when knockout cell line ab264817 (knockout cell lysate ab257544) was used. Wild-type and MVP knockout samples were subjected to SDS-PAGE. ab175239 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 2000 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-MVP antibody [EPR13227(B)] (ab175239) at 1/2000 dilution

Lane 1:

Wild-type HeLa lysate at 20 µg

Lane 2:

MVP knockout HeLa 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:

A549 lysate at 20 µg

Lane 4:

MOLT-4 lysate at 20 µg

Predicted band size: 99 kDa

Observed band size: 110 kDa

false

• WB

Lab

Western blot - Anti-MVP antibody [EPR13227(B)] (AB175239)

Blocking and dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-MVP antibody [EPR13227(B)] (ab175239) at 1/10000 dilution

Lane 1:

Calu-3 whole cell lysate at 20 µg

Lane 2:

A549 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: 53 kDa,55 kDa,73 kDa,97 kDa,99 kDa

Observed band size: 110 kDa

false

• WB

Lab

Western blot - Anti-MVP antibody [EPR13227(B)] (AB175239)

Blocking and dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-MVP antibody [EPR13227(B)] (ab175239) at 1/2000 dilution

All lanes:

Caco-2 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: 99 kDa

Observed band size: 110 kDa

false

• WB

CiteAb

Western blot - Anti-MVP antibody [EPR13227(B)] (AB175239)

MVP western blot using anti-MVP antibody [EPR13227(B)] ab175239. Publication image and figure legend from Shen, W., Qiu, Y., et al., 2019, Cancer Med, PubMed 31044552.

ab175239 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab175239 please see the product overview.

IL-25 regulates the NF-κB signaling pathway activity. A, The change of NF-κB signaling pathway in IL-25 overexpressed A549 cells and IL-25 silenced A549/CDDP cells. B, The effect of exogenous IL-25 on the activity of NF-κB signaling pathway in A549 cells. C, QNZ blocks the effect of exogenous IL-25 on the expression of major vault protein (MVP) and the activity of NF-κB signaling pathway in A549 cells. *p < 0.01 vs indicated controls, all figures represent the average of three sets of independent experiments

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