Anti-CACNA1E antibody [EPR25385-129]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CACNA1E antibody [EPR25385-129] (AB288766)

Positive staining on (A) HEK-293T cells transfected with a CACNA1E expression vector containing a his tag. No staining on (B) HEK-293T cells transfected with empty vector containing a his tag. The section was incubated with ab288766 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

Hematoxylin was used as a counter stain.

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CACNA1E antibody [EPR25385-129] (AB288766)

Negative control : No staining in rat kidney. The section was incubated with ab288766 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins was used. Hematoxylin was used as a counter stain.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CACNA1E antibody [EPR25385-129] (AB288766)

Positive staining in mouse striatum. The section was incubated with ab288766 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Counter stain used was Hematoxylin.

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CACNA1E antibody [EPR25385-129] (AB288766)

Immunohistochemical analysis of paraffin-embedded rat brain tissue labelling CACNA1E with ab288766 at 1/500 (0.956 μg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining in rat brain, and strong immunoreactivity was seen in the superficial layer of the cortex, hippocampus, and striatum. The section was incubated with ab288766 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

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-CACNA1E antibody [EPR25385-129] (AB288766)

Negative control : No staining in mouse kidney. The section was incubated with ab288766 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Hematoxylin was used as a counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) was used for 20 mins.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-CACNA1E antibody [EPR25385-129] (AB288766)

Rat primary neuron cell line was stained with primary ab288766 at 1/500 (Red). Rabbit monoclonal IgG (ab172730) (Black) was used as a isotype control at 1/2000. Cells were fixed in 4% paraformaldehyde and permeabilized with 90% methanol.

Secondary antibody used is Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at a 1/2000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CACNA1E antibody [EPR25385-129] (AB288766)

Positive staining in rat hippocampus. The section was incubated with ab288766 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

Hematoxylin was used as a counter stain.

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

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-CACNA1E antibody [EPR25385-129] (AB288766)

Mouse primary neuron cell line was stained with primary ab288766 at 1/500 (Red). Rabbit monoclonal IgG (ab172730) (Black) was used as a isotype control at 1/2000 dilution. Cells were fixed in 4% paraformaldehyde and permeabilized with 90% methanol.

Secondary antibody used is Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at a 1/2000 dilution.

• IP

Lab

Immunoprecipitation - Anti-CACNA1E antibody [EPR25385-129] (AB288766)

CACNA1E was immunoprecipitated from 0.35 mg mouse brain tissue lysate 10 μg with ab288766 at 1/30 dilution (2 μg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab288766 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : Mouse brain tissue lysate 10 μg

Lane 2 : ab288766 IP in mouse brain tissue lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab288766 in mouse brain tissue lysate

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

Exposure time : 67 seconds.

All lanes:

Immunoprecipitation - Anti-CACNA1E antibody [EPR25385-129] (ab288766)

Predicted band size: 262 kDa

Observed band size: 257 kDa

false

• WB

Lab

Western blot - Anti-CACNA1E antibody [EPR25385-129] (AB288766)

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

Lane 1 : 103 seconds
Lane 2-5 : 70 seconds

The molecular weight and expression pattern is consistent to what has been described in previous reports (PMID : 31704946, PMID : 30846929)

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

All lanes:

Western blot - Anti-CACNA1E antibody [EPR25385-129] (ab288766) 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 kidney tissue lysate at 40 µg

Lane 4:

Rat hippocampus tissue lysate at 40 µg

Lane 5:

Rat kidney tissue lysate at 40 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 1/100000 dilution

Predicted band size: 262 kDa

Observed band size: 257 kDa

false