Anti-Neurocan antibody [EPR23425-68]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neurocan antibody [EPR23425-68] (AB277525)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling Neurocan with ab277525 at 1/4000 (0.133 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human cerebrum (PMID : 9795216). The section was incubated with ab277525 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neurocan antibody [EPR23425-68] (AB277525)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling Neurocan with ab277525 at 1/4000 (0.133 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse cerebrum (PMID : 30356641) . The section was incubated with ab277525 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neurocan antibody [EPR23425-68] (AB277525)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling Neurocan with ab277525 at 1/4000 (0.133 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat cerebrum. The section was incubated with ab277525 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.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Neurocan antibody [EPR23425-68] (AB277525)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neuron cells labelling Neurocan with ab277525 at 1/50 (10.6 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in mouse primary neuron. Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection is observed. ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain MAP2 at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab277525 at 1/50 dilution, followed by ab150120 at 1/1000 dilution.

-ve control 2 : ab11267 at 1/200 dilution, followed by ab150077 at 1/1000 dilution.

• WB

Lab

Western blot - Anti-Neurocan antibody [EPR23425-68] (AB277525)

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

Neurocan is a glycoprotein. The observed MW is consistent with what has been described in the literature (PMID : 29670169).

Negative control : mouse skeletal muscle (PMID : 1326557)

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

Exposure time : 26 seconds.

All lanes:

Western blot - Anti-Neurocan antibody [EPR23425-68] (ab277525) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 40 µg

Lane 2:

Mouse hippocampus tissue lysate at 40 µg

Lane 3:

Mouse skeletal muscle tissue lysate at 40 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: 143 kDa

Observed band size: 130-300 kDa

false

• WB

Lab

Western blot - Anti-Neurocan antibody [EPR23425-68] (AB277525)

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

Neurocan is a glycoprotein. The observed MW is consistent with what has been described in the literature (PMID : 29670169).

Negative control : rat skeletal muscle (PMID : 1326557).

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

This blot was developed using a higher sensitivity ECL substrate.

Exposure time : 103 seconds.

All lanes:

Western blot - Anti-Neurocan antibody [EPR23425-68] (ab277525) at 1/1000 dilution

Lane 1:

Rat brain tissue lysate at 20 µg

Lane 2:

Rat hippocampus tissue lysate at 20 µg

Lane 3:

Rat skeletal muscle 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 1/100000 dilution

Predicted band size: 143 kDa

Observed band size: 130-300 kDa

false

• WB

Lab

Western blot - Anti-Neurocan antibody [EPR23425-68] (AB277525)

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

Neurocan proteolytic cleavage results in two fragments : neurocan-130 (130 kDa) and neurocan-C (150k Da ). Neurocan is a glycoprotein and seen as broad indistinct smears, which on chondroitinase treatment resolve into band of 150 (neurocan-C). The molecular weight observed is consistent with what has been described in the literature (PMID : 10729323).

Exposure time : 48 seconds.

All lanes:

Western blot - Anti-Neurocan antibody [EPR23425-68] (ab277525) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 15 µg

Lane 2:

Mouse brain tissue lysate treated with 0.05U chondroitinase ABC for 3h at 37℃ at 15 µg

Lane 3:

Rat brain tissue lysate at 15 µg

Lane 4:

Rat brain tissue lysate treated with 0.05U chondroitinase ABC for 3h at 37℃ at 15 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: 143 kDa

Observed band size: 150-300 kDa

false