Anti-DCC antibody [EPR23313-115]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DCC antibody [EPR23313-115] (AB273570)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling DCC with ab273570 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on human cerebrum (PMID : 11684721). The section was incubated with ab273570 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IP

Unknown

Immunoprecipitation - Anti-DCC antibody [EPR23313-115] (AB273570)

DCC was immunoprecipitated from 0.35 mg Human hippocampus tissue lysate with ab273570 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab273570 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Human hippocampus tissue lysate 10 ug

Lane 2 : ab273570 IP in Human hippocampus tissue lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab273570 in human hippocampus tissue lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 10 seconds.

All lanes:

Immunoprecipitation - Anti-DCC antibody [EPR23313-115] (ab273570)

Predicted band size: 158 kDa,176 kDa

Observed band size: 175 kDa,250 kDa

false

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DCC antibody [EPR23313-115] (AB273570)

Immunohistochemical analysis of paraffin-embedded Rat E14.5 tissue labeling DCC with ab273570 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on rat E14.5 cerebrum (PMID : 24739528). The section was incubated with ab273570 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• Flow Cyt

Unknown

Flow Cytometry - Anti-DCC antibody [EPR23313-115] (AB273570)

Flow cytometric analysis of PC-12 (Rat adrenal gland pheochromocytoma, Left) / Rat primary neuron cells (Right) cells labelling DCC with ab273570 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

Negative control : PC12 (PMID : 8044801).

Gated on viable cells.

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-DCC antibody [EPR23313-115] (AB273570)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat E14.5 embryo tissue labeling DCC with ab273570 at 1/100 dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (Green). Positive staining on rat embryonic brain is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• Flow Cyt

Unknown

Flow Cytometry - Anti-DCC antibody [EPR23313-115] (AB273570)

Flow cytometric analysis of Rat primary neuron cells cells labelling DCC with ab273570 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Black). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

Gated on viable cells.

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-DCC antibody [EPR23313-115] (AB273570)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse E14.5 embryo tissue labeling DCC with ab273570 at 1/100 dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (Green). Positive staining on mouse embryonic brain is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488)at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DCC antibody [EPR23313-115] (AB273570)

Immunohistochemical analysis of paraffin-embedded Mouse E14.5 tissue labeling DCC with ab273570 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on mouse E14.5 cerebrum (PMID : 24739528). The section was incubated with ab273570 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IP

Unknown

Immunoprecipitation - Anti-DCC antibody [EPR23313-115] (AB273570)

DCC was immunoprecipitated from 0.35 mg Mouse P5 cerebral cortex tissue lysate with ab273570 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab273570 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Mouse P5 cerebral cortex tissue lysate 10 ug

Lane 2 : ab273570 IP in Mouse P5 cerebral cortex tissue lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab273570 in Mouse P5 cerebral cortex tissue lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 32 seconds.

All lanes:

Immunoprecipitation - Anti-DCC antibody [EPR23313-115] (ab273570)

Predicted band size: 158 kDa

Observed band size: 175 kDa

false

• WB

Lab

Western blot - Anti-DCC antibody [EPR23313-115] (AB273570)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

DCC is a glycoprotein. The molecular weight observed is consistent with what have been described in the literature (PMID : 8044801).

Negative control : SW480 (PMID : 7926722).

Exposure time : 3 minutes.

All lanes:

Western blot - Anti-DCC antibody [EPR23313-115] (ab273570) at 1/1000 dilution

Lane 1:

Human hippocampus tissue lysate at 20 µg

Lane 2:

SW480 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

Mouse P5 hippocampus tissue lysate at 20 µg

Secondary

All lanes:

VeriBlot for IP secondary antibody(HRP)(<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Predicted band size: 158 kDa

Observed band size: 175 kDa

false

• WB

Lab

Western blot - Anti-DCC antibody [EPR23313-115] (AB273570)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

DCC is a glycoprotein. The molecular weight observed is consistent with what have been described in the literature (PMID : 8044801).

Samples are non-boiled as boiling may cause protein aggregates.

Exposure time : 26 seconds.

All lanes:

Western blot - Anti-DCC antibody [EPR23313-115] (ab273570) at 1/1000 dilution

Lane 1:

Rat P7 hippocampus tissue lysate at 20 µg

Lane 2:

Rat E18 brain tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 158 kDa

Observed band size: 175 kDa,240 kDa

false