Anti-Caspr antibody [K65/35] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspr antibody [K65/35] - BSA and Azide free (AB255761)

Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling Caspr with ab252535 at 1/10000 dilution, followed by ready to use secondary antibody. Positive staining on human cerebrum is observed. Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use secondary.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

The section was incubated with ab252535 for 10 mins at room temperature.

The immunostaining staining was performed on a Leica Biosystems BOND^® RX instrument.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252535).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Caspr antibody [K65/35] - BSA and Azide free (AB255761)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling Caspr with ab252535 at 1/50 dilution, followed by ab150113 AlexaFluor^®488 Goat anti-Mouse secondary antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in HeLa cell line. The nuclear counter stain is DAPI (Blue). Tubulin is detected using ab179504 Anti-beta IV Tubulin antibody - Microtubule Marker at 1/200 dilution, followed by ab150080 AlexaFluor^®594 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Red).

The negative controls are as follows :
-ve control 1 : ab252535 at 1/50 dilution, followed by ab150080 AlexaFluor^®594 Goat anti-Rabbit secondary at 1/1000 dilution.
-ve control 2 : ab179504 at 1/200 dilution, followed by ab150113 AlexaFluor^®488 Goat anti-Mouse secondary at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (252535).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Caspr antibody [K65/35] - BSA and Azide free (AB255761)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Neuro-2a (mouse neuroblastoma cell line) cells labeling Caspr with ab252535 at 1/50 dilution, followed by ab150113 AlexaFluor^®488 Goat anti-Mouse secondary antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in Neuro-2a cell line. The nuclear counter stain is DAPI (Blue). Tubulin is detected using ab179504 Anti-beta IV Tubulin antibody - Microtubule Marker at 1/200 dilution, followed by ab150080 AlexaFluor^®594 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Red).

The negative controls are as follows :
-ve control 1 : ab252535 at 1/50 dilution, followed by ab150080 AlexaFluor^®594 Goat anti-Rabbit secondary at 1/1000 dilution.
-ve control 2 : ab179504 at 1/200 dilution, followed by ab150113 AlexaFluor^®488 Goat anti-Mouse secondary at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (252535).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspr antibody [K65/35] - BSA and Azide free (AB255761)

Immunohistochemical analysis of paraffin-embedded rat cerebrum tissue labeling Caspr with ab252535 at 1/10000 dilution, followed by ready to use secondary antibody. Positive staining on rat cerebrum is observed. Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use secondary.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

The section was incubated with ab252535 for 10 mins at room temperature.

The immunostaining staining was performed on a Leica Biosystems BOND^® RX instrument.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252535).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspr antibody [K65/35] - BSA and Azide free (AB255761)

Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling Caspr with ab252535 at 1/10000 dilution, followed by ready to use secondary antibody. Positive staining on mouse cerebrum is observed. Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use secondary.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

The section was incubated with ab252535 for 10 mins at room temperature.

The immunostaining staining was performed on a Leica Biosystems BOND^® RX instrument.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252535).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Caspr antibody [K65/35] - BSA and Azide free (AB255761)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized C6 (rat glial tumor glial cell line) cells labeling Caspr with ab252535 at 1/50 dilution, followed by ab150113 AlexaFluor^®488 Goat anti-Mouse secondary antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in C6 cell line. The nuclear counter stain is DAPI (Blue). Tubulin is detected using ab179504 Anti-beta IV Tubulin antibody - Microtubule Marker at 1/200 dilution, followed by ab150080 AlexaFluor^®594 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Red).

The negative controls are as follows :
-ve control 1 : ab252535 at 1/50 dilution, followed by ab150080 AlexaFluor^®594 Goat anti-Rabbit secondary at 1/1000 dilution.
-ve control 2 : ab179504 at 1/200 dilution, followed by ab150113 AlexaFluor^®488 Goat anti-Mouse secondary at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (252535).

• WB

Supplier Data

Western blot - Anti-Caspr antibody [K65/35] - BSA and Azide free (AB255761)

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure time : 7.75 seconds.

Caspr protein (220 kd) was observed. The molecular weight is consistent with that has been described in the literature (PMID : 20610764).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (252535).

We recommend to use 1% SDS Hot lysis method to get desired WB results. For Lysate preparation protocol, please refer to the protocol book in the protocol section and/or here (downloadable copy).

All lanes:

Western blot - Anti-Caspr antibody [K65/35] (<a href='/ko/products/primary-antibodies/caspr-antibody-k65-35-ab252535'>ab252535</a>) at 1/1000 dilution

Lane 1:

Rat brain lysate at 20 µg

Lane 2:

Mouse brain lysate at 20 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Predicted band size: 156 kDa

Observed band size: 220 kDa

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