Anti-Integrin alpha V antibody [EPR19669] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin alpha V antibody [EPR19669] - BSA and Azide free (AB251483)

This data was developed using ab208012, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded human bladder cancer tissue labeling CD51 with ab208012 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Membrane staining on human bladder cancer is observed.

Counter stained with Hematoxylin.

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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin alpha V antibody [EPR19669] - BSA and Azide free (AB251483)

This data was developed using ab208012, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling CD51 with ab208012 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Cytoplasmic and membrane staining on human kidney is observed.

Counter stained with Hematoxylin.

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.

• WB

Lab

Western blot - Anti-Integrin alpha V antibody [EPR19669] - BSA and Azide free (AB251483)

This data was developed using ab208012, the same antibody clone in a different buffer formulation.

Western blot : Anti-Integrin alpha V antibody [EPR19669] ab208012 staining at 1/2000 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.

All lanes:

Western blot - Anti-Integrin alpha V antibody [EPR19669] (<a href='/en-us/products/primary-antibodies/integrin-alpha-v-antibody-epr19669-ab208012'>ab208012</a>) at 1/2000 dilution

Lane 1:

Wild-type HCT 116 at 20 µg

Lane 2:

Western blot - Human ITGAV knockout HCT116 cell line (<a href='/en-us/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

• WB

Supplier Data

Western blot - Anti-Integrin alpha V antibody [EPR19669] - BSA and Azide free (AB251483)

This data was developed using ab208012, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Integrin alpha V antibody [EPR19669] (<a href='/en-us/products/primary-antibodies/integrin-alpha-v-antibody-epr19669-ab208012'>ab208012</a>) at 1/2000 dilution

Lane 1:

Human fetal brain tissue lysate at 10 µg

Lane 2:

Human fetal heart tissue lysate at 10 µg

Lane 3:

Human fetal kidney tissue lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 116 kDa

Observed band size: 130-140 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-Integrin alpha V antibody [EPR19669] - BSA and Azide free (AB251483)

This data was developed using ab208012, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

The molecular weight observed is consistent with what has been described in the literature (PMID : 22740495).

All lanes:

Western blot - Anti-Integrin alpha V antibody [EPR19669] (<a href='/en-us/products/primary-antibodies/integrin-alpha-v-antibody-epr19669-ab208012'>ab208012</a>) at 1/2000 dilution

Lane 1:

A549 (Human lung carcinoma cell line) whole cell lysate at 10 µg

Lane 2:

A431 (Human epidermoid carcinoma cell line) whole cell lysate at 10 µg

Lane 3:

HT-29 (Human colorectal adenocarcinoma cell line) whole cell lysate at 10 µg

Lane 4:

HUVEC (Human umbilical vein endothelial 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: 116 kDa

Observed band size: 130-140 kDa

false

Exposure time: 5s

• WB

Supplier Data

Western blot - Anti-Integrin alpha V antibody [EPR19669] - BSA and Azide free (AB251483)

This data was developed using ab208012, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST. Exposure time : Lanes 1-3 : 5 seconds; Lanes 4 and 9 : 30 seconds; Lanes 5-7 : 1 minute; Lane 8 : 15 seconds.

All lanes:

Western blot - Anti-Integrin alpha V antibody [EPR19669] (<a href='/en-us/products/primary-antibodies/integrin-alpha-v-antibody-epr19669-ab208012'>ab208012</a>) at 1/2000 dilution

Lane 1:

C6 (Rat glial tumor cell line) whole cell lysate at 10 µg

Lane 2:

PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate at 10 µg

Lane 3:

NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate at 10 µg

Lane 4:

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

Lane 5:

Mouse brain tissue lysate at 10 µg

Lane 6:

Mouse kidney tissue lysate at 10 µg

Lane 7:

Mouse spleen tissue lysate at 10 µg

Lane 8:

Rat kidney tissue lysate at 10 µg

Lane 9:

Rat spleen tissue 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 100000 µg

Predicted band size: 116 kDa

Observed band size: 130-140 kDa

false