Anti-Presenilin 1/PS-1 antibody [APS 11] (ab15456)

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Overview

  • Product name

    Anti-Presenilin 1/PS-1 antibody [APS 11]
    See all Presenilin 1/PS-1 primary antibodies

  • Description

    Mouse monoclonal [APS 11] to Presenilin 1/PS-1

  • Host species

    Mouse

  • Specificity

    No cross-reactivity is seen with presenilin 2. In formalin-fixed, paraffin embedded sections of human brain, this antibody showed strong staining of both the plaque core and dystrophic neurites. By Western blot, this antibody detects an ~28 kDa protein representing PS1 N-terminus cleavage product in ST15 cell lysate transfected with human PS1.

  • Tested applications

    Suitable for: IP, IHC-FoFr, ICC, ICC/IF, WB, ELISA, IHC-P, Flow Cytmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human
    Predicted to work with: Cow, Dog, Non human primates, Cynomolgus monkey

  • Immunogen

    Synthetic peptide corresponding to Human Presenilin 1/PS-1 aa 21-34.
    Sequence:

    HLSNTVRSQNDNRE

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

  • Positive control

    • IHC: Human brain tissue slides. WB: ST15 cell lysate transfected with human PS1.

Properties

Applications

Our Abpromise guarantee covers the use of ab15456 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
IP Use at an assay dependent concentration. PubMed: 21163940
IHC-FoFr Use at an assay dependent concentration. PubMed: 18927449
ICC Use at an assay dependent concentration.
ICC/IF Use a concentration of 0.75 µg/ml.
WB Use a concentration of 12 µg/ml. Detects a band of approximately 28 kDa.
ELISA Use a concentration of 4.5 µg/ml.
IHC-P Use a concentration of 0.75 µg/ml.
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

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. Stimulates cell-cell adhesion though its association with the E-cadherin/catenin complex. Under conditions of apoptosis or calcium influx, cleaves E-cadherin promoting the disassembly of the E-cadherin/catenin complex and increasing the pool of cytoplasmic beta-catenin, thus negatively regulating Wnt signaling. May also play a role in hematopoiesis.

  • Tissue specificity

    Expressed in a wide range of tissues including various regions of the brain, liver, spleen and lymph nodes.

  • Involvement in disease

    Defects in PSEN1 are a cause of Alzheimer disease type 3 (AD3) [MIM:607822]. AD3 is a familial early-onset form of 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 PSEN1 are a cause of frontotemporal dementia [MIM:600274].
    Defects in PSEN1 are the cause of cardiomyopathy dilated type 1U (CMD1U) [MIM:613694]. 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.
    Defects in PSEN1 are the cause of acne inversa familial type 3 (ACNIF3) [MIM:613737]. A chronic relapsing inflammatory disease of the hair follicles characterized by recurrent draining sinuses, painful skin abscesses, and disfiguring scars. Manifestations typically appear after puberty.

  • 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 (NTF) and C-terminal (CTF) fragments of approximately 35 and 20 kDa, respectively. During apoptosis, the C-terminal fragment (CTF) is further cleaved by caspase-3 to produce the fragment, PS1-CTF12.
    After endoproteolysis, the C-terminal fragment (CTF) is phosphorylated on serine residues by PKA and/or PKC. Phosphorylation on Ser-346 inhibits endoproteolysis.

  • Cellular localization

    Endoplasmic reticulum membrane. Golgi apparatus membrane. Cell surface. Bound to NOTCH1 also at the cell surface. Colocalizes with CDH1/2 at sites of cell-cell contact. Colocalizes with CTNNB1 in the endoplasmic reticulum and the proximity of the plasma membrane. Also present in azurophil granules of neutrophils.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • AD3 antibody
    • Ad3h antibody
    • FAD antibody
    • Homo Sapiens Clone CC44 Senilin 1 antibody
    • Presenilin-1 CTF12 antibody
    • Protein S182 antibody
    • PS 1 antibody
    • PS-1 antibody
    • PS1-CTF12 antibody
    • PSEN1 antibody
    • PSN1_HUMAN antibody
    • PSNL1 antibody
    • S182 antibody
    see all

Images

  • Immunocytochemistry/ Immunofluorescence - Anti-Presenilin 1/PS-1 antibody [APS 11] (ab15456)
    Immunocytochemistry/ Immunofluorescence - Anti-Presenilin 1/PS-1 antibody [APS 11] (ab15456)
    IF staining PS1 using ab15456.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Presenilin 1/PS-1 antibody [APS 11] (ab15456)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Presenilin 1/PS-1 antibody [APS 11] (ab15456)
    Immunohistochemistry was performed on normal biopsies of deparaffinized Human liver tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:200 with a mouse monoclonal antibody recognizing Presenilin 1 ab15456 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Presenilin 1/PS-1 antibody [APS 11] (ab15456)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Presenilin 1/PS-1 antibody [APS 11] (ab15456)
    Immunohistochemistry was performed on normal biopsies of deparaffinized Human brain tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:20 with a mouse monoclonal antibody recognizing Presenilin 1 ab15456 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

  • Flow Cytometry - Anti-Presenilin 1/PS-1 antibody [APS 11] (ab15456)
    Flow Cytometry - Anti-Presenilin 1/PS-1 antibody [APS 11] (ab15456)
    Overlay histogram showing HepG2 cells stained with ab15456 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab15456, 1ug/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG1 (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was Mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HepG2 cells fixed with 4% paraformaldehyde (10 min) permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

References

This product has been referenced in:

  • Zoltowska KM  et al. Dynamic presenilin 1 and synaptotagmin 1 interaction modulates exocytosis and amyloid ß production. Mol Neurodegener 12:15 (2017). Read more (PubMed: 28193235) »
  • Raven F  et al. Soluble Gamma-secretase Modulators Attenuate Alzheimer's ß-amyloid Pathology and Induce Conformational Changes in Presenilin 1. EBioMedicine 24:93-101 (2017). Read more (PubMed: 28919280) »
See all 8 Publications for this product

Publishing research using ab15456? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-7 of 7 Abreviews or Q&A

Question

So we use fresh dissected hippocampus from rat brain, its immediately placed in lysis buffer, homogenzied, and stored at -80. We got clear bands for actin, as well as a number of other proteins for these samples. For this antibody specifically, we block in 5% milk/TBS, incubate the primary overnight at -4 degrees. We have used 1:1000, and 1:250 concentrations of the antibody with no bands. We then incubate for 1hr at RT with a licor secondary, wash with TBST and image. This protocol generally works with minor tweaking for most antibodies. Whats strange is getting zero band imaging with the licor, usually at least something shows up.

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Answer

Thank you for your reply.


Based on the information you have provided, I would recommend increasing the amount of antibody used in the WB. The datasheet calls for 12ug/ml for WB and you have only used up to 4ug/ml. I would recommend increasing the concentration to observe optimal performance from this antibody.

I hope this information is helpful. Please do not hesitate to contact us if you have any additional questions.

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Question

I was wondering if you had any references for Anti-Presenilin 1 antibody [APS 11] (ab15456) used in rat brain samples?

We are attempting to use this antibody, and have followed the recommended guidelines, but are not seeing any bands at 28kda.

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Answer

According to our records, ab15456 was proving difficult to use in WB and we were in contact in order to help resolve the issue.

Looking at our correspondence, it appears that we are awaiting more details in order to help us better understand the difficulties experienced. If the requested information has already been sent, it appears that it did not reach our Scientific Support team and we apologize for this inconvenience. In this case we would like to ask for the information again so that we can reach a resolution.

If the issue has already been settled, please let us know so that we can be assured that the problem has been solved to your satisfaction and update our records.

We wish you the best of luck with your research and look forward to a reply.

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Question

I was wondering if you had any references for Anti-Presenilin 1 antibody [APS 11] (ab15456) used in rat brain samples? We are attempting to use this antibody, and have followed the recommended guidelines, but are not seeing any bands at 28kda.

Read More
Answer

Thank you for contacting Abcam regarding ab15456.


I am sorry that you are experiencing difficulties with this antibody in WB. I have checked our available references, and unfortunately none specifically use rat brains as samples.


I would be happy to review your protocol information and provide some suggestions. Please provide a detailed protocol summary and sample preparation information. If I am unable to make any suggestions to improve your results, I am happy to resolve this issue per our Abpromise guarantee.


I look forward to your reply so that I may assist you further. Please do not hesitate to contact me if you have any additional questions.

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Question

I have been using one of your antibodies, ab15456 Anti-Presenilin 1, and I was just woundering if you have a control peptide that you use for blocking this antibody in immunofluorescence, to use as a control for specific antibody staining. Do you have a peptide like that?

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Answer

Thank you for your enquiry. I'm afraid we do not have the immunizing peptide to use for blocking control experiments for ab15456. I'm very sorry I cannot help you more,

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Question

Thank you for your quick reply. We would like to test one more of your antibody as a replacement for ab15456, and found that ab15458 could be a good replacement. We also looked for a N-terminal PS1 antibody but did not find any that was tested against Hum, rat and mouse. Do you have any? Hope that this will turn out OK now.

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Answer

Let's see - we have the following PS1 antibodies that have been tested for cross-reactivity with human, mouse, and rat and tested in Western blotting: ab15456 (which you tried), ab15458, ab16244 (a new chicken polyclonal), and ab10281 (tested in human, predicted to cross-react with mouse and rat due to sequence homology). So just let me know which you would like. Also, what was the Abcam order number or purchase order number that was used for your order for ab15456? Thank you!

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Question

I have now done exactly what you recommended me to do. Unfortunately I got a very unclean blot with unspecific bands. Is it possible to get any other PS1 antibody in test size from you? We are getting desperate in our looks after a good PS1 antibody. Best Regards

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Answer

Thank you for your email and I'm sorry to hear that you are still experiencing difficulty with ab15456. I'm replying to you on behalf of Sarah, who is away from the office this week. As Sarah mentioned, we can send you a replacement antibody or offer you a refund. We do have several other Presenilin 1 antibodies. To see the list, go to www.abcam.com and type in Presenilin 1 in the "search for" box located at the upper left hand side of the page. Some suggestions are ab16244 and ab15458 - both have been tested in Western blotting and for cross-reactivity with rat.

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Question

BATCH NUMBER 77220 ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM Non-specific band I have SHSY5Y cells gives 1 band around 40kDa ,fibroblast no bands,rat liver ~ 70 kDa, rat brain no bands. With a higher Konc. of the Ab I get alot of unspecific bands SAMPLE SHSY5Y cell lysate, MEFPS1Wt cell Lysate ,Rat liver homogenate, rat brain homogenaite and BD8 cell lysate as an negative control. All at an conc. of 25?g/sample PRIMARY ANTIBODY abcam/ab15456/ 12?g/ml in TBS-T with 5%non fat milk /over night in room temp./wash 4x15min in TBS-T before Secondary Ab SECONDARY ANTIBODY Amersham/NA931V ,Anti-mouseIgG. Horseradish Peroxidase linked whole Ab (from Sheep) /1:2000 DETECTION METHOD Pierce, SuperSignal West Pico Chemiluminiscent Substrate Prod# 34080 POSITIVE AND NEGATIVE CONTROLS USED I have used the BD8 cells as negativ control The SHSY5Y as positiv. ANTIBODY STORAGE CONDITIONS I diluted the send batch in aliquots and stored them in -80 degree C. I used 12?g/ml in TBS-T with 5%non fat milk when I inkubaited my memebrains. SAMPLE PREPARATION The cells are lysed in TBS with NP40 and triton X-100 The homogenate are made in an Mannitol/Hepes buffer. The samles are diluted in 2x Laemmi samplebuffer They are not heated but I let them stay for at least 20min in sample buffer before i put them on the gel. AMOUNT OF PROTEIN LOADED 25?g ELECTROPHORESIS/GEL CONDITIONS 10-20% creterion gel TRANSFER AND BLOCKING CONDITIONS Transfer buffer:Tris+Glysin. This transferbuffer we all use and it is working fine Transfer 55-60min acording to the Criterion protocol.The marker transfers nice. HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? ~ 10 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? I have chanced the concentration of the first Ab. ADDITIONAL NOTES We really need a PS1 and also a PS2 Ab that works we have. Right now we don?t have any good. What we would like, is to get a small amount of different PS1 and PS2 to test, because it is getting really expensive to by Ab that is not working. Hope for help.

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Answer

I'm very sorry you have a problem with ab15456, thank you for taking the time to answer our questionnaire, it is very useful. I would like to suggest the following modifications to your protocol: -use the recommended positive control of human brain lysate -use protease inhibitors in your lysis buffer (preferably a lysis buffer like the RIPA buffer) -following addition of the loading buffer to your samples, boil at 95-100C for 5min, so that the protein is denatured and reduced and can be recognised by the antibody -block in BSA 5% in TBST 1hr -incubate the primary antibody in TBST (no blocking agent) overnight at 4C. Please be reassured that should you still have a problem after these changes we will offer you a replacement or credit note. I hope this advice helps, please let us know how you get on,

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