Anti-Doublecortin antibody

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Doublecortin antibody (AB235153)

Human brain tissue stained for Doublecortin using ab235153 at 1/1000 dilution in immunohistochemical analysis. The counter stain is hematoxylin.

Heat induced epitope retrieval (HIER).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Doublecortin antibody (AB235153)

4% PFA fixed, 0.3% Triton X-100 permeabilized post-natal rat pup (PND1) heterogeneous brain cells labeling Doublecortin using ab235153 at 15 μg/ml (O/N at 4°C) (green) in ICC/IF. The Secondary antibody is a Donkey anti-Gt IgG DyLigh™ 488 conjugated preadsorbed, used at 5 μg/ml for 1 hour at room temperature. The Nuclear counterstain is DAPI (Blue).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Doublecortin antibody (AB235153)

Formalin-fixed, paraffin-embedded mouse dentate gyrus tissue stained for Doublecortin with ab235153 at a 1/200 dilution (30 mins at room temperature) in immunohistochemical analysis. A Donkey anti Goat (HRP) secondary was used at 4 μl/ml (20 mins at room temperature). Counterstained with hematoxylin. HIER using citrate buffer for 20 mins.

Pathologist analysis : Doublecortin (DCX) showed selective staining of soma and nuclei of cells in the dentate gyrus in mouse brain. The specificity of the staining is consistent with DCX staining in the Human Protein Atlas and as such, is suitable for IHC at a concentration of 1/200.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Doublecortin antibody (AB235153)

4% PFA fixed, 0.3% Triton X-100 permeabilized post-natal rat pup (PND1) heterogeneous brain cells labeling Doublecortin using ab235153 at 15 μg/ml (O/N at 4°C) (green) in ICC/IF.The Secondary antibody is a Donkey anti-Gt IgG DyLigh™ 488 conjugated preadsorbed antibody, used at 5 μg/ml for 1 hour at room temperature. The Nuclear counterstain is DAPI (Blue).

• WB

Supplier Data

Western blot - Anti-Doublecortin antibody (AB235153)

* "Doublecortin microtubule affinity is regulated by a balance of kinase and phosphatase activity at the leading edge of migrating neurons." Schaar B.T., Kinoshita K., McConnell S.K. Neuron 41 : 203213 (2004).

Minimal cortex defined : small portion of cortex removed prior to lysis.

All lanes:

Western blot - Anti-Doublecortin antibody (ab235153) at 1/1000 dilution

Lane 1:

HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) cell lysate at 15 µg

Lane 2:

Post-natal rat pup whole brain lysate (PND2-6)* at 15 µg

Lane 3:

Rat pup PND2-6 - minimal cortex brain lysate at 15 µg

Lane 4:

Rat pup PND2-6 – minimal cortex brain lysate at 35 µg

Secondary

All lanes:

Western blot - Donkey Anti-Goat IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/donkey-goat-igg-h-l-hrp-preadsorbed-ab7125'>ab7125</a>) at 1/40000 dilution

Predicted band size: 49 kDa

Observed band size: 45 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-Doublecortin antibody (AB235153)

All lanes:

Western blot - Anti-Doublecortin antibody (ab235153) at 1/1000 dilution

Lane 1:

Doublecortin (human) overexpressing HEK-293T lysate C-Myc/DDK tagged

Lane 2:

HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) cell lysate

Lane 3:

Adult mouse normal brain tissue lysate

Lane 4:

Adult rat normal brain tissue lysate

Secondary

All lanes:

Western blot - Donkey Anti-Goat IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/donkey-goat-igg-h-l-hrp-preadsorbed-ab7125'>ab7125</a>) at 1/40000 dilution

Predicted band size: 49 kDa

Observed band size: 35 kDa,48 kDa

false

Exposure time: 15s