Anti-Caspr antibody [EPR7828]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspr antibody [EPR7828] (AB133634)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cerebrum tissue sections labeling Caspr with Purified ab133634 at 1 : 2000 dilution (0.29 µg/mL). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspr antibody [EPR7828] (AB133634)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human astrocytoma tissue sections labeling Caspr with Purified ab133634 at 1 : 2000 dilution (0.29 µg/mL). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Caspr antibody [EPR7828] (AB133634)

Immunocytochemistry/ Immunofluorescence analysis of Neuro-2a (mouse neuroblastoma neuroblast) cells labeling Caspr with purified ab133634 at 1/100 (5.7 μg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1/200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/1000 (2 μg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• WB

Lab

Western blot - Anti-Caspr antibody [EPR7828] (AB133634)

Lanes 1 - 4 : Merged signal (red and green). Green - ab133634 observed at 175 kDa. Red - loading control, ab18058, observed at 130 kDa.

ab133634 was shown to recognize Caspr in wild-type HAP1 cells as signal was lost at the expected MW in CNTNAP1 (Caspr) knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and CNTNAP1 (Caspr) knockout samples were subjected to SDS-PAGE. ab133634 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Caspr antibody [EPR7828] (ab133634) at 1/1000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

CNTNAP1 (Caspr) knockout HAP1 whole cell lysate at 20 µg

Lane 3:

MEF whole cell lysate at 20 µg

Lane 4:

HeLa whole cell lysate at 20 µg

Predicted band size: 156 kDa

false

• WB

Unknown

Western blot - Anti-Caspr antibody [EPR7828] (AB133634)

The molecular weight is consistent with that has been described in the literature (PMID : 20610764).

We are unsure about the nature of extra bands.

All lanes:

Western blot - Anti-Caspr antibody [EPR7828] (ab133634) at 1/1000 dilution

Lane 1:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate boiled at 20 µg

Lane 2:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate unboiled at 20 µg

Lane 3:

SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate boiled at 20 µg

Lane 4:

SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate unboiled at 20 µg

Lane 5:

Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysate boiled at 20 µg

Lane 6:

Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysate unboiled at 20 µg

Lane 7:

PC-12 (Rat adrenal gland pheochromocytoma ) whole cell lysate boiled at 20 µg

Lane 8:

PC-12 (Rat adrenal gland pheochromocytoma ) whole cell lysate unboiled at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 156 kDa

Observed band size: 220 kDa

false