Anti-Integrin alpha V antibody [EPR16800] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Integrin alpha V antibody [EPR16800] - BSA and Azide free (AB271932)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% Triton X-100 permeabilized A549 (Human lung carcinoma) cells labeling Integrin alpha V with ab179475 at 1/500 dilution, followed by Goat anti-rabbit Alexa Fluor^® 488 (IgG) (ab150077) secondary antibody at 1/400 dilution (green).

Confocal image showing membrane and cytoplasm staining on A549 cell line. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and ab150120 (goat anti-mouse AlexaFluor^®594 secondary antibody) at 1/500 dilution (red).

The negative controls are as follows :

-ve control 1 : ab179475 at 1/500 dilution followed by ab150120 (AlexaFluor^®594 Goat anti-Mouse secondary) at 1/500 dilution.

-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Alexa Fluor^®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179475).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin alpha V antibody [EPR16800] - BSA and Azide free (AB271932)

Immunohistochemical analysis of paraffin-embedded human transitional cell carcinoma of bladder tissue labeling Integrin alpha V with ab179475 at 1/500 dilution followed by Goat Anti-Rabbit HRP (IgG H&L) (ab97051) secondary antibody at 1/500 dilution.

Membrane and weak cytoplasm staining on human transitional cell carcinoma of bladder is observed. Counterstained with hematoxylin.

Negative control : Using 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179475).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin alpha V antibody [EPR16800] - BSA and Azide free (AB271932)

Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling Integrin alpha V with ab179475 at 1/500 dilution followed by Goat Anti-Rabbit HRP (IgG H&L) (ab97051) secondary antibody at 1/500 dilution.

Membrane and cytoplasm staining on human kidney tubules is observed. Counterstained with hematoxylin.

Negative control : Using 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179475).

• IP

Supplier Data

Immunoprecipitation - Anti-Integrin alpha V antibody [EPR16800] - BSA and Azide free (AB271932)

Integrin alpha V was immunoprecipitated from 1 mg of A549 (Human lung carcinoma) whole cell extract with ab179475 at 1/40 dilution. Western blot analysis was performed from the immunoprecipitate using ab179475 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

Lane 1 : A549 whole cell extract.

Lane 2 : PBS instead of A549 whole cell extract.

Blocking/Dilution buffer : 5% NFDM/TBST.

ab179475 can recognize 135kDa full length Integrin alpha V and 125kDa heavy chain. The 125 kDa band is Integrin alpha V heavy chain.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179475).

All lanes:

Immunoprecipitation - Anti-Integrin alpha V antibody [EPR16800] (<a href='/ko/products/primary-antibodies/integrin-alpha-v-antibody-epr16800-ab179475'>ab179475</a>)

Predicted band size: 116 kDa

Observed band size: 125 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin alpha V antibody [EPR16800] - BSA and Azide free (AB271932)

Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling Integrin alpha V with ab179475 at 1/500 dilution followed by Goat Anti-Rabbit HRP (IgG H&L) (ab97051) secondary at 1/500 dilution.

Membrane and cytoplasm staining on mouse kidney tubule and glomerulus is observed. Counterstained with hematoxylin.

Negative control : Using 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179475).

• WB

Lab

Western blot - Anti-Integrin alpha V antibody [EPR16800] - BSA and Azide free (AB271932)

This data was developed using ab179475, the same antibody clone in a different buffer formulation.

Western blot : Anti-Integrin alpha V antibody [EPR16800] ab179475 staining at 1/5000 dilution, shown in green; Mouse anti alpha Tubulin ab7291 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 116 kDa in Wild-type HCT 116 cell lysates with no signal observed at this size in ITGAV knockout HCT 116 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 100pc LICOR in TBS-0.1 % Tween 20 (TBS-T) before incubation with primary antibodies overnight at 4 C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

Lanes 1 - 4:

Western blot - Anti-Integrin alpha V antibody [EPR16800] (<a href='/ko/products/primary-antibodies/integrin-alpha-v-antibody-epr16800-ab179475'>ab179475</a>) at 1/5000 dilution

Lanes 1 - 4:

Western blot - Anti-Integrin alpha V antibody [EPR16800] - Low endotoxin, Azide free (<a href='/ko/products/primary-antibodies/integrin-alpha-v-antibody-epr16800-low-endotoxin-azide-free-ab222222'>ab222222</a>) at 1/5000 dilution

Lanes 1 - 4:

Western blot - Anti-Integrin alpha V antibody [EPR16800] - BSA and Azide free (Detector) (<a href='/ko/products/primary-antibodies/integrin-alpha-v-antibody-epr16800-bsa-and-azide-free-detector-ab245051'>ab245051</a>) at 1/5000 dilution

Lane 1:

Wild-type HCT 116 at 20 µg

Lane 2:

Western blot - Human ITGAV knockout HCT116 cell line (<a href='/ko/products/cell-lines/human-itgav-knockout-hct116-cell-line-ab286702'>ab286702</a>) at 20 µg

Lane 3:

A549 at 20 µg

Lane 4:

Daudi at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 116 kDa

Observed band size: 116 kDa

false