Anti-Presenilin 2/AD5 antibody [EP1515Y] - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Presenilin 2/AD5 antibody [EP1515Y] - BSA and Azide free (AB239841)

This data was developed using ab51249, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded human Alzheimer's brain tissue* labelling PSEN2 with ab51249 at 1ug/ml followed by a ready to use LeicaDS9800 (Bond^™ Polymer Refine Detection).

Positive staining in human Alzheimer's brain.

The section was incubated with ab51249 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 sodium citrate buffer (pH 6.0 epitope retrieval solution1) for 20 mins.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Presenilin 2/AD5 antibody [EP1515Y] - BSA and Azide free (AB239841)

Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Presenilin 2/AD5 with purified ab51249 at 1/50 dilution (5μg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab51249).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Presenilin 2/AD5 antibody [EP1515Y] - BSA and Azide free (AB239841)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cerebrum tissue sections labeling Presenilin 2/AD5 with purified ab51249 at 1/500 dilution (0.51 μg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab51249).

• IP

Unknown

Immunoprecipitation - Anti-Presenilin 2/AD5 antibody [EP1515Y] - BSA and Azide free (AB239841)

ab51249 (purified ) at 1/20 dilution (1ug) immunoprecipitating Presenilin 2/AD5 in HeLa whole cell lysate.

Lane 1 (input) : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10ug
Lane 2 (+) : ab51249 & HeLa whole cell lysate
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab51249 in HeLa whole cell lysate

For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.

Blocking and diluting buffer : 5% NFDM/TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab51249).

All lanes:

Immunoprecipitation - Anti-Presenilin 2/AD5 antibody [EP1515Y] (<a href='/en-us/products/primary-antibodies/presenilin-2-ad5-antibody-ep1515y-ab51249'>ab51249</a>)

Predicted band size: 50 kDa

false

• IP

Supplier Data

Immunoprecipitation - Anti-Presenilin 2/AD5 antibody [EP1515Y] - BSA and Azide free (AB239841)

Presenilin was immunoprecipitated from 1 mg of HeLa whole cell extract with ab51249 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab51249 at 1/5000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

Input : HeLa whole cell extract 10 μg.
IP (+) : ab51249 IP in HeLa whole cell extract.
IP (-) : Rabbit monoclonal IgG (ab172730) instead of ab51249 in HeLa whole cell extract.

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab51249).

All lanes:

Immunoprecipitation - Anti-Presenilin 2/AD5 antibody [EP1515Y] (<a href='/en-us/products/primary-antibodies/presenilin-2-ad5-antibody-ep1515y-ab51249'>ab51249</a>)

Predicted band size: 50 kDa

Observed band size: 24 kDa

false

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Presenilin 2/AD5 antibody [EP1515Y] - BSA and Azide free (AB239841)

Immunocytochemistry/ Immunofluorescence analysis of PC-12 (Rat adrenal gland pheochromocytoma) cells labeling Presenilin 2/AD5 with purified ab51249 at 1/50 dilution (5 μg/ml). Cells were fixed in 100% Methanol and permeabilized with None. 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab51249).

• IP

Unknown

Immunoprecipitation - Anti-Presenilin 2/AD5 antibody [EP1515Y] - BSA and Azide free (AB239841)

Presenilin was immunoprecipitated from 1 mg of PC-12 (rat adrenal gland pheochromocytoma) whole cell extract with ab51249 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab51249 at 1/5000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

Input : PC-12 whole cell extract 10 μg.
IP (+) : ab51249 IP in PC-12 whole cell extract.
IP (-) : Rabbit monoclonal IgG (ab172730) instead of ab51249 in PC-12 whole cell extract.

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab51249).

All lanes:

Immunoprecipitation - Anti-Presenilin 2/AD5 antibody [EP1515Y] (<a href='/en-us/products/primary-antibodies/presenilin-2-ad5-antibody-ep1515y-ab51249'>ab51249</a>)

Predicted band size: 50 kDa

Observed band size: 24 kDa

false

• WB

Lab

Western blot - Anti-Presenilin 2/AD5 antibody [EP1515Y] - BSA and Azide free (AB239841)

This data was developed using ab51249, the same antibody clone in a different buffer formulation.

Western blot : Rabbit Monoclonal[EP1515Y] to Presenilin 2/AD5 ab51249 staining at 1/20000 dilution, shown in green; Mouse anti GAPDH (ab8245) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 18 kDa in Wild-type U-87 MG ab278079 cell lysates with no signal observed at this size in PSEN2 knockout U-87 MG cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-Presenilin 2/AD5 antibody [EP1515Y] (<a href='/en-us/products/primary-antibodies/presenilin-2-ad5-antibody-ep1515y-ab51249'>ab51249</a>) at 1/20000 dilution

Lane 1:

Wild-type U-87 MG ab278079 at 20 µg

Lane 2:

PSEN2 knockout U-87 MG at 20 µg

Lane 3:

A549 at 20 µg

Lane 4:

MOLT-4 at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 50 kDa

Observed band size: 18 kDa

false