Anti-Integrin alpha 2 antibody [2/CD49b]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin alpha 2 antibody [2/CD49b] (AB300628)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Integrin alpha 2 with ab300628 at 1/2000 (0.516 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on human colon.The section was incubated with ab300628 for 30 mins at room temperature, followed by anti-mouse IgG2a antibody for 8 mins during the LeicaDS9800 kit staining procedure. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin alpha 2 antibody [2/CD49b] (AB300628)

Immunohistochemical analysis of paraffin-embedded Human cervical cancer tissue labeling Integrin alpha 2 with ab300628 at 1/2000 (0.516 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on human cervical cancer.The section was incubated with ab300628 for 30 mins at room temperature, followed by anti-mouse IgG2a antibody for 8 mins during the LeicaDS9800 kit staining procedure. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• WB

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Western blot - Anti-Integrin alpha 2 antibody [2/CD49b] (AB300628)

Blocking and diluting buffer and concentration : Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBSFalse colour image of Western blot : Anti-Integrin alpha 2 antibody [2/CD49b] (ab300628) staining at 1/1000 dilution, shown in green; Rabbit anti-GAPDH antibody [16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab300628 was shown to bind specifically to ITGA2. A band was observed at 150 kDa in wild-type HeLa cell lysates with no signal observed at this size in the ITGA2 knockout cell line. To generate this image, wild-type and ITGA2 knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a PVDF-FL membrane. Membranes were blocked in Odyssey diluted in equal volume of 0.1 % TBS 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-Mouse IgG H&L (IRDye® 800CW) (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) (ab216777) at 1/10000 dilution. Performed under reducing conditions. Exposure time :

All lanes:

Western blot - Anti-Integrin alpha 2 antibody [2/CD49b] (ab300628) at 1/1000 dilution

Lane 1:

Wild-type HeLa (human cervix adenocarcinoma epithelial cell) cell lysate

Lane 2:

ITGA2 knockout HeLa (human cervix adenocarcinoma epithelial cell) cell lysate

Lane 3:

A431 (human epidermoid carcinoma epithelial cell) cell lysate

Secondary

Lanes 1 - 3:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-800cw-preadsorbed-ab216772'>ab216772</a>) at 1/10000 dilution

Lanes 1 - 3:

Western blot - Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-680rd-preadsorbed-ab216777'>ab216777</a>) at 1/10000 dilution

false

• WB

Supplier Data

Western blot - Anti-Integrin alpha 2 antibody [2/CD49b] (AB300628)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Exposure time : Lanes 1-2 : 26 seconds Lane 3 : 15 seconds

All lanes:

Western blot - Anti-Integrin alpha 2 antibody [2/CD49b] (ab300628) at 1/1000 dilution

Lane 1:

A549 (human lu carcinoma epithelial cell), whole cell lysate 10

Lane 2:

293T (human embryonic kidney epithelial cell), whole cell lysate

Lane 3:

U-2 OS (human bone osteosarcoma epithelial cell), whole cell lysate

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

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