Anti-CASK antibody [PT55]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CASK antibody [PT55] (AB326349)

Immunocytochemistry/Immunofluorescence analysis. Tissue : Neuroblastoma cells (Kelly). Species : Human. Fixation : 4% PFA for 10 min at RT. Permeabilization : 0.15% TritonX for 15 min at RT. Blocking : 10% goat serum for 40 min at RT. ab326349 at 1 : 100 for 1 hour at RT. Secondary Antibody : Goat anti-Rabbit IgG-AlexaFluor488 (green) at 1 : 1000 for 1 hour at RT in the dark. Counterstain : DAPI (blue) nuclear stain at 1 : 1000 for 5 min at RT in the dark. Note : Phalloidin stain was used to visualize any potential nuclear F-actin.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CASK antibody [PT55] (AB326349)

Immunohistochemistry analysis. Tissue : Brain. Species : Mouse. Fixation : Paraffin embedded, 4um thin. Blocking : 3% milk for 1 hour at RT. Primary Antibody : ab326349 at 1 : 100 for 1 hour at RT. Secondary Antibody : Goat anti-rabbit IgG-HRP (H&L) at 1 : 100 for 45 min at RT. Counterstain : Hematoxylin (purple) nuclear stain at 1 : 10 for 5 min at RT. Color Development : DAB at 1 : 50 for 15 min at RT.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CASK antibody [PT55] (AB326349)

Immunocytochemistry/Immunofluorescence analysis. Tissue : Pheochromocytoma cells (PC-12). Species : Rat. Fixation : 4% PFA for 10 min at RT. Permeabilization : 0.15% TritonX for 15 min at RT. Blocking : 10% goat serum for 40 min at RT. Primary Antibody : ab326349 at 1 : 100 for 1 hour at RT. Secondary Antibody : Goat anti-Rabbit IgG-AlexaFluor488 (green) at 1 : 1000 for 1 hour at RT in the dark. Counterstain : DAPI (blue) nuclear stain at 1 : 1000 for 5 min at RT in the dark.

• WB

Supplier Data

Western blot - Anti-CASK antibody [PT55] (AB326349)

Western blot analysis showing detection of Human CASK protein in a rat brain membrane lysate. Block : 2% skim milk + 2% BSA for 1 hour at RT. Primary Antibody : ab326349 at 1 : 1000 for 2 hours at RT. Secondary Antibody : Goat anti-rabbit IgG-HRP (H&L) at 1 : 5000 for 1 hour at RT. Color Development : Chemiluminescent for HRP for 3 min at RT in the dark. Exposed 1 second.

All lanes:

Western blot - Anti-CASK antibody [PT55] (ab326349) at 1/1000 dilution

All lanes:

rat brain membrane lysate at 10 µg

Secondary

All lanes:

Goat anti-rabbit IgG-HRP (H&L) at 1/5000 dilution

false

Exposure time: 1s

• WB

Supplier Data

Western blot - Anti-CASK antibody [PT55] (AB326349)

Western blot analysis showing detection of Human CASK protein in a transient over-expression (OE) lysate. Block : 2% skim milk + 2% BSA for 1 hour at RT. Primary Antibody : ab326349 at 1 : 10,000 for 2 hours at RT. Secondary Antibody : Goat anti-rabbit IgG-HRP (H&L) at 1 : 5000 for 1 hour at RT. Color Development : Chemiluminescent for HRP for 1 min at RT in the dark. Exposed 1 second. NOTE : The ORF clone contains a C-term DDK tag; results were confirmed using Anti-DDK antibody, showing band at ~45kDa is related to a C-term fragment of the CASK protein (data not shown). Literature explains the presence of the ~70kDa by stating that phosphorylation of CASK can lead to its degradation which may produce multiple fragments, including a 70kDa fragment.

All lanes:

Western blot - Anti-CASK antibody [PT55] (ab326349) at 1/10000 dilution

Lane 1:

HEK293T OE Lysate at 2.5 µg

Lane 2:

HEK293T Neg. Ctrl at 2.5 µg

Secondary

All lanes:

Goat anti-rabbit IgG-HRP (H&L) at 1/5000 dilution

false

Exposure time: 1s