Overview

  • Product name

    Anti-Presenilin 2/AD5 antibody [EP1515Y]
    See all Presenilin 2/AD5 primary antibodies
  • Description

    Rabbit monoclonal [EP1515Y] to Presenilin 2/AD5
  • Host species

    Rabbit
  • Specificity

    The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cytmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Presenilin 2/AD5 aa 300-400 (C terminal). The exact sequence is proprietary.

  • Positive control

    • WB: HeLa, PC-12, HepG2, C2C12, C6, and Neuro-2a lysates. IHC-P: human cerebrum and kidney tissues. ICC/IF: PC-12, HepG2, JAR and A-673 cells. FC: HeLa cells. IP: HeLa cells.
  • General notes

    Previously labelled as Presenilin 2. 

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

Applications

Our Abpromise guarantee covers the use of ab51249 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/20000. Detects a band of approximately 23 kDa (predicted molecular weight: 50 kDa).
IHC-P 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

ICC/IF 1/50 - 1/250.
IP 1/20.
Flow Cyt 1/50.

Target

  • Function

    Probable catalytic subunit of the gamma-secretase complex, an endoprotease complex that catalyzes the intramembrane cleavage of integral membrane proteins such as Notch receptors and APP (beta-amyloid precursor protein). Requires the other members of the gamma-secretase complex to have a protease activity. May play a role in intracellular signaling and gene expression or in linking chromatin to the nuclear membrane. May function in the cytoplasmic partitioning of proteins.
  • Tissue specificity

    Isoform 1 is seen in the placenta, skeletal muscle and heart while isoform 2 is seen in the heart, brain, placenta, liver, skeletal muscle and kidney.
  • Involvement in disease

    Defects in PSEN2 are the cause of Alzheimer disease type 4 (AD4) [MIM:606889]. AD is an autosomal dominant Alzheimer disease. Alzheimer disease is a neurodegenerative disorder characterized by progressive dementia, loss of cognitive abilities, and deposition of fibrillar amyloid proteins as intraneuronal neurofibrillary tangles, extracellular amyloid plaques and vascular amyloid deposits. The major constituent of these plaques is the neurotoxic amyloid-beta-APP 40-42 peptide (s), derived proteolytically from the transmembrane precursor protein APP by sequential secretase processing. The cytotoxic C-terminal fragments (CTFs) and the caspase-cleaved products such as C31 derived from APP, are also implicated in neuronal death.
    Defects in PSEN2 are the cause of cardiomyopathy dilated type 1V (CMD1V) [MIM:613697]. It is a disorder characterized by ventricular dilation and impaired systolic function, resulting in congestive heart failure and arrhythmia. Patients are at risk of premature death.
  • Sequence similarities

    Belongs to the peptidase A22A family.
  • Domain

    The PAL motif is required for normal active site conformation.
  • Post-translational
    modifications

    Heterogeneous proteolytic processing generates N-terminal and C-terminal fragments.
    Phosphorylated on serine residues.
  • Cellular localization

    Endoplasmic reticulum membrane. Golgi apparatus membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • AD3L antibody
    • AD3LP antibody
    • AD4 antibody
    • AD5 antibody
    • Alzheimer disease 4 antibody
    • CMD1V antibody
    • E5-1 antibody
    • OTTHUMP00000035671 antibody
    • OTTHUMP00000035672 antibody
    • OTTHUMP00000228286 antibody
    • OTTHUMP00000228288 antibody
    • Presenilin 2 (Alzheimer disease 4) antibody
    • Presenilin 2 antibody
    • Presenilin-2 CTF subunit antibody
    • PS-2 antibody
    • PS2 antibody
    • Psen2 antibody
    • PSN2_HUMAN antibody
    • PSNL2 antibody
    • STM-2 antibody
    • STM2 antibody
    see all

Images

  • 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� 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.
  • Anti-Presenilin 2/AD5 antibody [EP1515Y] (ab51249) at 1/1000 dilution ((unpurified) for 16 hours at 4°C) + HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 30 µg

    Secondary
    HRP-conjugated Goat anti-rabbit IgG at 1/2000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 50 kDa
    Observed band size: 24 kDa why is the actual band size different from the predicted?


    Exposure time: 1 minute


    Blocking Step: 5% Milk for 1 hour at 24°C

    See Abreview

  • Immunofluorescent analysis of 100% methanol-fixed, 0.1% Triton X-100 permeabilized A-673 (Human muscle Ewing's Sarcoma cell line) cells labeling Presenilin with unpurified ab51249 at 1/250 dilution, followed by Anti-rabbit Alexa Fluor® 488 (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing cytoplasmic and weakly nuclear staining on A-673 cells. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and Anti-Mouse AlexaFluor® 594 (ab150120) at 1/500 dilution (red).
    The negative controls are as follows:
    -ve control 1: ab51249 at 1/250 dilution followed by Anti-Mouse AlexaFluor® 594 (ab150120) at 1/500 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by Anti-Rabbit Alexa Fluor® 488 (ab150077) at 1/500 dilution.

     

  • 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.

     

  • Anti-Presenilin 2/AD5 antibody [EP1515Y] (ab51249) at 1/200000 dilution ((unpuriifed)) + PC-12 (rat adrenal gland pheochromocytoma cell line) cell lysate at 10 µg

    Secondary
    Goat anti-rabbit HRP labeled at 1/2000 dilution

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

  • Unpurified ab51249 at a 1/50 dilution staining Presenilin 2/AD5 in human kidney tissue by immunohistochemistry using paraffin embedded tissue.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Immunofluorescent analysis of 100% methanol-fixed, 0.1% Triton X-100 permeabilized JAR (human placenta choriocarcinoma cell line) cells labeling Presenilin with unpurified ab51249 at 1/250 dilution, followed by Anti-rabbit Alexa Fluor® 488 (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing cytoplasm staining on JAR cells. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and Anti-Mouse AlexaFluor® 594 (ab150120) at 1/500 dilution (red).
    The negative controls are as follows:
    -ve control 1: ab51249 at 1/250 dilution followed by Anti-Mouse AlexaFluor® 594 (ab150120) at 1/500 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by Anti-Rabbit Alexa Fluor® 488 (ab150077) at 1/500 dilution.

  • 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.

     

References

This product has been referenced in:

  • Braggin JE  et al. Alternative splicing in a presenilin 2 variant associated with Alzheimer disease. Ann Clin Transl Neurol 6:762-777 (2019). Read more (PubMed: 31020001) »
  • Liu L  et al. A cellular complex of BACE1 and ?-secretase sequentially generates Aß from its full-length precursor. J Cell Biol 218:644-663 (2019). Read more (PubMed: 30626721) »
See all 9 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Sodium Citrate, 0.05% Tween 20, pH 6.0
Permeabilization
No
Specification
brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
Paraformaldehyde

Sulieman Minhas

Verified customer

Submitted Dec 13 2016

Application
Western blot
Sample
Human Cell lysate - whole cell (human iPS cell derived neurons (healthy/diseased))
Gel Running Conditions
Reduced Denaturing (12.5% SDS gel)
Loading amount
10 µg
Specification
human iPS cell derived neurons (healthy/diseased)
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: rt°C

Abcam user community

Verified customer

Submitted Jan 25 2016

Question
Answer

Thank you for contacting us.

The immunogen is located within residues 300 - 350 for this antibody.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Read More
Application
Western blot
Sample
Mouse Cell lysate - other (WT AND KO MEF)
Gel Running Conditions
Reduced Denaturing
Loading amount
30 µg
Specification
WT AND KO MEF
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

Abcam user community

Verified customer

Submitted Nov 08 2010

Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (HELA)
Total protein in input
200 µg
Immuno-precipitation step
Protein G
Specification
HELA

Abcam user community

Verified customer

Submitted Sep 22 2010

Application
Immunocytochemistry
Sample
Human Cultured Cells (HeLa)
Permeabilization
Yes - Triton-X-0.5%
Specification
HeLa
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 24°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Sep 14 2010

Application
Western blot
Sample
Human Cell lysate - whole cell (HELA)
Gel Running Conditions
Reduced Denaturing
Loading amount
30 µg
Specification
HELA
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

Abcam user community

Verified customer

Submitted Aug 30 2010

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