Phospho-Doublecortin (Ser47) Polyclonal Antibody

$Paraformaldehyde-fixed, paraffin embedded Rat brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with Doublecortin (Ser47) Polyclonal Antibody, Unconjugated (bs-3113R) at 1:400 overnight at 4\u00b0C, DAB staining.$

Paraformaldehyde-fixed, paraffin embedded Rat brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with Doublecortin (Ser47) Polyclonal Antibody, Unconjugated (bs-3113R) at 1:400 overnight at 4\u00b0C, DAB staining.

U251 cell lysates probed with Doublecortin (Ser47) Polyclonal Antibody, Unconjugated (bs-3113R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.

Human HepG2 Cells were fixed with 2% PFA (10min at room temperature) and then permeabilized with 0.2% PBST for 30min at room temperature. The cells were then incubated in 2% BSA + 10% goat serum to block non-specific protein-protein interactions for 15 min at room temperature. Cells were stained with Doublecortin (Ser47) Polyclonal Antibody, Unconjugated (bs-3113R) at 1:100 for 30 min at room temperature. The secondary antibody, Goat anti-rabbit IgG-FITC (bs-0295G-FITC), was used for 40 min at room temperature. Primary Antibody staining (green) is compared to unstained cells (black), secondary only (blue) and isotype control (bs-0295P; orange).

Lane 1: Mouse Cerebrum lysates; Lane 2: Rat Cerebrum lysates probed with Doublecortin (Ser47) Polyclonal Antibody, Unconjugated (bs-3113R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.

$SHSY5Y cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37\u00b0C for 20 min; Antibody incubation with (Phospho-Doublecortin (Ser128)) polyclonal Antibody, Unconjugated (bs-3113R) 1:100, 90 minutes at 37\u00b0C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37\u00b0C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.$

SHSY5Y cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37\u00b0C for 20 min; Antibody incubation with (Phospho-Doublecortin (Ser128)) polyclonal Antibody, Unconjugated (bs-3113R) 1:100, 90 minutes at 37\u00b0C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37\u00b0C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.

Applications

WB
ELISA
FCM
IHC-P
IHC-F
IF(IHC-P)
IF(IHC-F)
IF(ICC)

Reactivity

Human
Mouse
Rat

Predicted Reactivity

Dog
Cow
Horse
Chicken

Overview
Catalog #	bs-3113R
Product Name	Phospho-Doublecortin (Ser47) Polyclonal Antibody
Applications	WB ELISA FCM IHC-P IHC-F IF(IHC-P) IF(IHC-F) IF(ICC)
Reactivity	Human, Mouse, Rat
Predicted Reactivity	Dog, Cow, Horse, Chicken
Specifications
Conjugation	Unconjugated
Host	Rabbit
Source	KLH conjugated synthetic phosphopeptide derived from human Doublecortin around the phosphorylation site of Ser47
Modification Site	Ser47
Clonality	Polyclonal
Isotype	IgG
Concentration	1ug/ul
Purification	Purified by Protein A.
Storage Buffer	0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
Storage Condition	Shipped at 4_. Store at -20_ for one year. Avoid repeated freeze/thaw cycles.
Target
Gene ID	1641
Swiss Prot	O43602
Subcellular location	Cytoplasm
Synonyms	Doublecortex; DBCN; Dbct; DC; Doublin; Lis X; Lissencephalin X; Lissencephaly X linked; Lissencephaly X linked doublecortin; LISX; Neuronal migration protein doublecortin; SCLH; XLIS; p-DCX; p-Doublecortex; phospho-Doublecortex.
Background	Neuronal Marker Doublecortin (DCX) is a microtubule-associated protein expressed almost exclusively in immature neurons. Neuronal precursors begin to express DCX shortly after exiting the cell cycle, and continue to express DCX for 2-3 weeks as the cells mature into neurons. Downregulation of DCX begins after 2 weeks, and occurs at the same time that these cells begin to express, a marker for mature neurons. Due to the nearly exclusive expression of DCX in developing neurons, this protein has been used increasingly as a marker for neurogenesis. Indeed, the levels of DCX expression increase in response to exercise, which occurs in parallel with increased BrdU labelling, currently a "gold standard" in measuring neurogenesis.
Application Dilution
WB	1:300-5000
ELISA	1:500-1000
FCM	1:20-100
IHC-P	1:200-400
IHC-F	1:100-500
IF(IHC-P)	1:50-200
IF(IHC-F)	1:50-200
IF(ICC)	1:50-200