Anti-alpha Adaptin抗体[AC1-M11]

• WB

Supplier Data

Western blot - Anti-alpha Adaptin antibody [AC1-M11] (AB2807)

Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10 % Bis-Tris gel, XCell SureLock™ Electrophoresis System and Novex® Sharp Pre-Stained Protein Standard. Resolved proteins were then transferred onto a nitrocellulose membrane with Pierce™ Power Blotter System (22834). The membrane was probed with the relevant primary and secondary Antibody using iBind™ Flex Western Starter Kit. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate.

All lanes:

Western blot - Anti-alpha Adaptin antibody [AC1-M11] (ab2807) at 2 µg/mL

Lane 1:

Rat brain tissue lysate at 30 µg

Lane 2:

SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate at 30 µg

Lane 3:

U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) whole cell lysate at 30 µg

Lane 4:

COLO 205 (Human colon adenocarcinoma cell line) whole cell lysate at 30 µg

Lane 5:

PC-3 (Human prostate adenocarcinoma cell line) whole cell lysate at 30 µg

Lane 6:

MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 30 µg

Lane 7:

NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate at 30 µg

Lane 8:

A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 30 µg

Secondary

All lanes:

Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate at 1/2500 dilution

Predicted band size: 108 kDa

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• 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.

• ICC/IF

Supplier Data

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

Immunolocalization of alpha-adaptin in NRK cells using ab2807.

• 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 (low power image of image 2).