Anti-Integrin alpha 2 antibody [EPR17338] - BSA and Azide free

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Integrin alpha 2 antibody [EPR17338] - BSA and Azide free (AB271936)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181548).

Flow cytometry overlay histogram showing staining with ab181548 of HAP1 WT positive cells (magenta line) and HAP1-ITGA2 KO negative cells (red line). The cells were fixed with 80% methanol, and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab181548) (1x 10⁶ in 100μl at 0.2μg/ml (1/2,500)) for 30min at 22°C.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C.

Isotype control antibody was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control in HAP1 WT cells (black line) and HAP1-ITGA2 KO cells (grey line), used at the same concentration and conditions as the primary antibody.

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Integrin alpha 2 antibody [EPR17338] - BSA and Azide free (AB271936)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized PC-3 (Human prostate adenocarcinoma cell line) cells labeling integrin alpha 2 with ab181548 at 1/100 dilution, followed by Goat anti-rabbit IAlexa Fluor® 488 (IgG) (ab150077) secondary antibody at 1/400 dilution (green). Confocal image showing membrane and weakly cytoplasmic staining on PC-3 cell line is observed. 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 - ab181548 at 1/100 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 (ab181548).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin alpha 2 antibody [EPR17338] - BSA and Azide free (AB271936)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Integrin alpha 2 with ab181548 at 1/500 dilution followed by Goat Anti-Rabbit HRP (IgG H&L) (ab97051) at 1/500 dilution. Membrane and weak cytoplasmic staining on epithelial cells of human colon is observed. Counter stained with Hematoxylin.

Negative control : Used PBS instead of primary antibody.

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 (ab181548).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Integrin alpha 2 antibody [EPR17338] - BSA and Azide free (AB271936)

ab181548 staining Integrin α2 in wild-type HAP1 cells (top panel) and Integrin α2 knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab181548 at 1μg/ml concentration and ab7291 at 1ug/ml concentration overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to Mouse IgG (Alexa Fluor® 594) (ab150120) at 2ug/ml (shown in pseudo-color red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181548).

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Integrin alpha 2 antibody [EPR17338] - BSA and Azide free (AB271936)

Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed A549 (Human lung carcinoma) cells labeling integrin alpha 2 with ab181549 at 1/160 dilution (red) compared with a rabbit monoclonal IgG isotype control (black) and a unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181548).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Integrin alpha 2 antibody [EPR17338] - BSA and Azide free (AB271936)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MCF7 (Human breast adenocarcinoma cell line) cells labeling integrin alpha 2 with ab181548 at 1/100 dilution, followed by Goat anti-rabbit IAlexa Fluor® 488 (IgG) (ab150077) secondary antibody at 1/400 dilution (green). Confocal image showing membrane staining on MCF7 cell line is observed. 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 - ab181548 at 1/100 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 (ab181548).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin alpha 2 antibody [EPR17338] - BSA and Azide free (AB271936)

Immunohistochemical analysis of paraffin-embedded Human squamous cell carcinoma of cervix tissue labeling Integrin alpha 2 with ab181548 at 1/500 dilution followed by Goat Anti-Rabbit HRP (IgG H&L) (ab97051) at 1/500 dilution. Membrane and weak cytoplasmic staining on epithelial cells of human squamous cell carcinoma of cervix tissue is observed. Counter stained with Hematoxylin.
Negative control : Used PBS instead of primary antibody.

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 (ab181548).

• IP

Supplier Data

Immunoprecipitation - Anti-Integrin alpha 2 antibody [EPR17338] - BSA and Azide free (AB271936)

Integrin alpha 2 was immunoprecipitated from 1mg of T-47D (Human ductal breast epithelial tumor cell line) whole cell extract with ab181548 at 1/150 dilution. Western blot was performed using ab181548 at 1/20,000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution. Lane 1 : T-47D whole cell extract Lane 2 : PBS instead of T-47D whole cell extract.

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181548).

All lanes:

Immunoprecipitation - Anti-Integrin alpha 2 antibody [EPR17338] - C-terminal (<a href='/en-us/products/primary-antibodies/integrin-alpha-2-antibody-epr17338-c-terminal-ab181548'>ab181548</a>)

Predicted band size: 129 kDa

Observed band size: 150 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin alpha 2 antibody [EPR17338] - BSA and Azide free (AB271936)

Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling Integrin alpha 2 with ab181548 at 1/500 dilution followed by Goat Anti-Rabbit HRP (IgG H&L) (ab97051) at 1/500 dilution. Membrane staining on epithelial cells of Rat colon tissue is observed. Counter stained with Hematoxylin.
Negative control : Used PBS instead of primary antibody.

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 (ab181548).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin alpha 2 antibody [EPR17338] - BSA and Azide free (AB271936)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling Integrin alpha 2 with ab181548 at 1/500 dilution followed by Goat Anti-Rabbit HRP (IgG H&L) (ab97051) at 1/500 dilution. Membrane and weak cytoplasmic staining on epithelial cells of Mouse kidney tubule is observed. Counter stained with Hematoxylin.
Negative control : Used PBS instead of primary antibody.

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 (ab181548).