All lanes : Anti-Presenilin 2/AD5 antibody [EP1515Y] (ab51249) at 1/20000 dilution (Purified)

Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
Lane 2 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates
Lane 3 : C2C12 (Mouse myoblasts myoblast) whole cell lysates
Lane 4 : C6 (Rat glial tumor glial cell) whole cell lysates
Lane 5 : Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysates

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 50 kDa
Observed band size: 23 kDa why is the actual band size different from the predicted?

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.

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.

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).

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.

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.

 

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.