Anti-alpha Adaptin antibody [AC1-M11]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha Adaptin antibody [AC1-M11] (AB2807)

Immunohistochemistry was performed on normal biopsies of deparaffinized Human heart tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer and microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1 : 20 with a Mouse monoclonal antibody recognizing alpha Adaptin (ab2807) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

• Flow Cyt

Unknown

Flow Cytometry - Anti-alpha Adaptin antibody [AC1-M11] (AB2807)

Overlay histogram showing HepG2 cells stained with ab2807 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab2807, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 2μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HepG2 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-alpha Adaptin antibody [AC1-M11] (AB2807)

Immunolocalization of alpha-adaptin in NRK cells using ab2807.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-alpha Adaptin antibody [AC1-M11] (AB2807)

Immunolocalization of alpha-adaptin in NRK cells using ab2807 (low power image of image 2).

• ICC/IF

CiteAb

Immunocytochemistry/ Immunofluorescence - Anti-alpha Adaptin antibody [AC1-M11] (AB2807)

Immunocytochemistry-immunofluorescence using Anti-alpha Adaptin antibody [AC1-M11], ab2807. Publication image from Poincloux, R. et al., 2018, Nat Commun, 30237420. Legend direct from paper.

αvβv5 integrin localizes to plaques and is required for their assembly. a HeLa cells were seeded on collagen-coated glass and fixed 24 h later before being stained forα-adaptin andαvβ5-integrin. Scale bar : 10 µm. Higher magnifications of boxed regions are shown. Arrows point to clathrin-coated plaques; arrowheads point to CCPs. b Quantification ofαvβ5 enrichment at plaques versus CCPs (*P < 0.005, two tailed Student’s t-test. n = 3; 100 structures per experiment were counted.). c Quantification of colocalization (Pearson’s coefficient) betweenαvβ5 andα-adaptin in cells cultured on the indicated collagen-coated substrate (*P < 0.005; **P < 0.001, one-way analysis of variance—ANOVA. n = 3). d HeLa cells treated with control (upper panel) or β5-specific (lower panel) siRNAs were seeded on collagen-coated glass and fixed 24 h later before being stained forα-adaptin. Scale bar : 15 µm. e Kymographs showing CCS dynamics in genome-edited HeLa cells treated with the indicated siRNA, seeded on collagen-coated glass, and imaged by spinning disk microscopy every 5 s for 5 min. f Quantification of the dynamics of CCSs observed as in e and treated with the indicated siRNAs (**P < 0.001, ANOVA. n = 3). g HeLa cells treated with β5-specific siRNAs were transfected with a siRNA-resistant β 5-GFP encoding construct and then fixed 24 h later before being stained forα-adaptin. Scale bar : 10 µm. The star marks a cell that is not transfected by β5-GFP. Higher magnifications of boxed regions are shown. All results are expressed as mean ± SD

• ICC/IF

CiteAb

Immunocytochemistry/ Immunofluorescence - Anti-alpha Adaptin antibody [AC1-M11] (AB2807)

Immunocytochemistry-immunofluorescence using Anti-alpha Adaptin antibody [AC1-M11], ab2807. Publication image from Poincloux, R. et al., 2018, Nat Commun, 30237420. Legend direct from paper.

αvβv5 integrin localizes to plaques and is required for their assembly. a HeLa cells were seeded on collagen-coated glass and fixed 24 h later before being stained forα-adaptin andαvβ5-integrin. Scale bar : 10 µm. Higher magnifications of boxed regions are shown. Arrows point to clathrin-coated plaques; arrowheads point to CCPs. b Quantification ofαvβ5 enrichment at plaques versus CCPs (*P < 0.005, two tailed Student’s t-test. n = 3; 100 structures per experiment were counted.). c Quantification of colocalization (Pearson’s coefficient) betweenαvβ5 andα-adaptin in cells cultured on the indicated collagen-coated substrate (*P < 0.005; **P < 0.001, one-way analysis of variance—ANOVA. n = 3). d HeLa cells treated with control (upper panel) or β5-specific (lower panel) siRNAs were seeded on collagen-coated glass and fixed 24 h later before being stained forα-adaptin. Scale bar : 15 µm. e Kymographs showing CCS dynamics in genome-edited HeLa cells treated with the indicated siRNA, seeded on collagen-coated glass, and imaged by spinning disk microscopy every 5 s for 5 min. f Quantification of the dynamics of CCSs observed as in e and treated with the indicated siRNAs (**P < 0.001, ANOVA. n = 3). g HeLa cells treated with β5-specific siRNAs were transfected with a siRNA-resistant β 5-GFP encoding construct and then fixed 24 h later before being stained forα-adaptin. Scale bar : 10 µm. The star marks a cell that is not transfected by β5-GFP. Higher magnifications of boxed regions are shown. All results are expressed as mean ± SD

• ICC/IF

CiteAb

Immunocytochemistry/ Immunofluorescence - Anti-alpha Adaptin antibody [AC1-M11] (AB2807)

Immunocytochemistry-immunofluorescence using Anti-alpha Adaptin antibody [AC1-M11], ab2807. Publication image from Poincloux, R. et al., 2018, Nat Commun, 30237420. Legend direct from paper.

αvβv5 integrin localizes to plaques and is required for their assembly. a HeLa cells were seeded on collagen-coated glass and fixed 24 h later before being stained forα-adaptin andαvβ5-integrin. Scale bar : 10 µm. Higher magnifications of boxed regions are shown. Arrows point to clathrin-coated plaques; arrowheads point to CCPs. b Quantification ofαvβ5 enrichment at plaques versus CCPs (*P < 0.005, two tailed Student’s t-test. n = 3; 100 structures per experiment were counted.). c Quantification of colocalization (Pearson’s coefficient) betweenαvβ5 andα-adaptin in cells cultured on the indicated collagen-coated substrate (*P < 0.005; **P < 0.001, one-way analysis of variance—ANOVA. n = 3). d HeLa cells treated with control (upper panel) or β5-specific (lower panel) siRNAs were seeded on collagen-coated glass and fixed 24 h later before being stained forα-adaptin. Scale bar : 15 µm. e Kymographs showing CCS dynamics in genome-edited HeLa cells treated with the indicated siRNA, seeded on collagen-coated glass, and imaged by spinning disk microscopy every 5 s for 5 min. f Quantification of the dynamics of CCSs observed as in e and treated with the indicated siRNAs (**P < 0.001, ANOVA. n = 3). g HeLa cells treated with β5-specific siRNAs were transfected with a siRNA-resistant β 5-GFP encoding construct and then fixed 24 h later before being stained forα-adaptin. Scale bar : 10 µm. The star marks a cell that is not transfected by β5-GFP. Higher magnifications of boxed regions are shown. All results are expressed as mean ± SD