Anti-BACE2 antibody [EPR23339-221] - BSA and Azide free

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BACE2 antibody [EPR23339-221] - BSA and Azide free (AB270616)

Immunohistochemical analysis of paraffin-embedded mouse pancreas tissue labeling BACE2 with ab270458 at 1/100 followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on mouse pancreatic islet (PMID : 21907142). The section was incubated with ab270458 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab270458).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-BACE2 antibody [EPR23339-221] - BSA and Azide free (AB270616)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% Triton X-100 permeabilized Beta-TC-6 cells labeling BACE2 with ab270458 at 1/50 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplamic and membranous staining in Beta-TC-6 cell line. 100% methanol fixation is recommended. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) 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 (ab270458).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BACE2 antibody [EPR23339-221] - BSA and Azide free (AB270616)

Immunohistochemical analysis of paraffin-embedded rat pancreas tissue labeling BACE2 with ab270458 at 1/100 followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on rat pancreatic islet (PMID : 21907142). The section was incubated with ab270458 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab270458).

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-BACE2 antibody [EPR23339-221] - BSA and Azide free (AB270616)

Immunohistochemical analysis of 4% PFA fixed 0.2% Triton X-100 permeabilized frozen mouse pancreas tissue labeling BACE2 with ab270458 at 1/100 dilution followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 (2 μg/ml) dilution. The nuclear counterstain was DAPI (Blue).

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

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody was ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 (2 μg/ml) dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab270458).

• IP

Unknown

Immunoprecipitation - Anti-BACE2 antibody [EPR23339-221] - BSA and Azide free (AB270616)

BACE2 was immunoprecipitated from 0.35 mg Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate with ab270458 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab270458 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Neuro-2a whole cell lysate.

Lane 2 : ab270458 IP in Neuro-2a whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab270458 in Neuro-2a whole cell lysate.

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

Exposure time : 3 mins.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab270458).

All lanes:

Immunoprecipitation - Anti-BACE2 antibody [EPR23339-221] (<a href='/en-us/products/primary-antibodies/bace2-antibody-epr23339-221-ab270458'>ab270458</a>)

Predicted band size: 56 kDa

false

• WB

Supplier Data

Western blot - Anti-BACE2 antibody [EPR23339-221] - BSA and Azide free (AB270616)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab270458). Blocking and diluting buffer and concentration : 5% NFDM/TBST. Brain has very low level of BACE2 [PMID : 10749877, PMID : 30626751].

All lanes:

Western blot - Anti-BACE2 antibody [EPR23339-221] (<a href='/en-us/products/primary-antibodies/bace2-antibody-epr23339-221-ab270458'>ab270458</a>) at 1/1000 dilution

Lane 1:

Beta-TC-6 (Mouse pancreas insulinoma beta cell) whole cell lysate at 20 µg

Lane 2:

Mouse brain tissue lysate at 20 µg

Lane 3:

Mouse hippocampus tissue lysate at 20 µg

Lane 4:

Mouse cerebellum tissue lysate at 20 µg

Lane 5:

Mouse cerebral cortex tissue lysate at 20 µg

Lane 6:

Mouse spinal cord tissue lysate at 20 µg

Lane 7:

Mouse midbrain tissue lysate at 20 µg

Lane 8:

Mouse forebrain tissue lysate 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: 56 kDa

Observed band size: 56 kDa

false