Anti-Presenilin 2/AD5 antibody [EP1515Y]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Presenilin 2/AD5 antibody [EP1515Y] (AB51249)

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] (AB51249)

Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Presenilin 2/AD5 with purified ab51249 at 1/50 dilution (5 μg/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).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Presenilin 2/AD5 antibody [EP1515Y] (AB51249)

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^TM 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.

• IP

Unknown

Immunoprecipitation - Anti-Presenilin 2/AD5 antibody [EP1515Y] (AB51249)

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.

All lanes:

Immunoprecipitation - Anti-Presenilin 2/AD5 antibody [EP1515Y] (ab51249)

Predicted band size: 50 kDa

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• IP

Supplier Data

Immunoprecipitation - Anti-Presenilin 2/AD5 antibody [EP1515Y] (AB51249)

Presenilin was immunoprecipitated from 1 mg of HeLa whole cell extract with unpurified 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.

All lanes:

Immunoprecipitation - Anti-Presenilin 2/AD5 antibody [EP1515Y] (ab51249)

Predicted band size: 50 kDa

Observed band size: 24 kDa

false

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Presenilin 2/AD5 antibody [EP1515Y] (AB51249)

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.

• IP

Unknown

Immunoprecipitation - Anti-Presenilin 2/AD5 antibody [EP1515Y] (AB51249)

Presenilin was immunoprecipitated from 1 mg of PC-12 (rat adrenal gland pheochromocytoma) whole cell extract with unpurified 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.

All lanes:

Immunoprecipitation - Anti-Presenilin 2/AD5 antibody [EP1515Y] (ab51249)

Predicted band size: 50 kDa

Observed band size: 24 kDa

false

• WB

Lab

Western blot - Anti-Presenilin 2/AD5 antibody [EP1515Y] (AB51249)

All lanes:

Western blot - Anti-Presenilin 2/AD5 antibody [EP1515Y] (ab51249) at 1/20000 dilution

Lane 1:

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

Lane 2:

HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates at 20 µg

Lane 3:

C2C12 (Mouse myoblasts myoblast) whole cell lysates at 20 µg

Lane 4:

C6 (Rat glial tumor glial cell) whole cell lysates at 20 µg

Lane 5:

Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysates 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: 50 kDa

Observed band size: 23 kDa

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• WB

Lab

Western blot - Anti-Presenilin 2/AD5 antibody [EP1515Y] (AB51249)

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] (ab51249) 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