Home Primary Antibodies Anti-Presenilin 1/PS-1 Rabbit Antibody [G24D17] For research use only.

Anti-Presenilin 1/PS-1 Rabbit Antibody [G24D17]

Catalog No.: F3873

    Application: Reactivity:

    Usage Information

    Dilution
    1:1000 - 1:5000
    1:100
    Application
    WB, IHC
    Reactivity
    Mouse, Rat, Human
    Source
    Rabbit
    Storage Buffer
    PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3
    Storage (from the date of receipt)
    -20°C (avoid freeze-thaw cycles), 2 years
    Predicted MW Observed MW
    53 kDa 18 kDa
    *Why do the predicted and actual molecular weights differ?
    The following reasons may explain differences between the predicted and actual protein molecular weight.
    Positive Control Mouse brain; Rat brain; Human fetal brain; Human cerebral cortex; Human Alzheimer's brain; Human glioma; HAP1; HEK293; HeLa; PC-3
    Negative Control

    Exprimental Methods

    WB
    Experimental Protocol:
     
    Sample preparation
    1. Tissue: Lyse the tissue sample by adding an appropriate volume of ice-cold RIPA/NP-40 Lysis Buffer (containing Protease Inhibitor Cocktail)，and homogenize the tissue at a low temperature.
    2. Adherent cell: Aspirate the culture medium and wash the cells with ice-cold PBS twice. Lyse the cells by adding an appropriate volume of RIPA/NP-40 Lysis Buffer (containing Protease Inhibitor Cocktail) and put the sample on ice for 5 min.
    3. Suspension cell: Transfer the culture medium to a pre-cooled centrifuge tube. Centrifuge and aspirate the supernatant. Wash the cells with ice-cold PBS twice. Lyse the cells by adding an appropriate volume of RIPA/NP-40 Lysis Buffer (containing Protease Inhibitor Cocktail) and put the sample on ice for 5 min.
    4. Place the lysate into a pre-cooled microcentrifuge tube. Centrifuge at 4°C for 15 min. Collect the supernatant;
    5. Remove a small volume of lysate to determine the protein concentration;
    6. Combine the lysate with protein loading buffer. Boil 20 µL sample under 95-100°C for 5 min. Centrifuge for 5 min after cool down on ice.
     
    Electrophoretic separation
    1. According to the concentration of extracted protein, load appropriate amount of protein sample and marker onto SDS-PAGE gels for electrophoresis. Recommended separating gel (lower gel) concentration: 10%. Reference Table for Selecting SDS-PAGE Separation Gel Concentrations
    2. Power up 80V for 30 minutes. Then the power supply is adjusted (110 V~150 V), the Marker is observed, and the electrophoresis can be stopped when the indicator band of the predyed protein Marker where the protein is located is properly separated. (Note that the current should not be too large when electrophoresis, too large current (more than 150 mA) will cause the temperature to rise, affecting the result of running glue. If high currents cannot be avoided, an ice bath can be used to cool the bath.)
     
    Transfer membrane
    1. Take out the converter, soak the clip and consumables in the pre-cooled converter;
    2. Activate PVDF membrane with methanol for 1 min and rinse with transfer buffer;
    3. Install it in the order of "black edge of clip - sponge - filter paper - filter paper - glue -PVDF membrane - filter paper - filter paper - sponge - white edge of clip";
    4. The protein was electrotransferred to PVDF membrane. ( 0.45 µm PVDF membrane is recommended ) Reference Table for Selecting PVDF Membrane Pore Size Specifications
    Recommended conditions for wet transfer: 200 mA, 120 min.
    ( Note that the transfer conditions can be adjusted according to the protein size. For high-molecular-weight proteins, a higher current and longer transfer time are recommended. However, ensure that the transfer tank remains at a low temperature to prevent gel melting.)
     
    Block
    1. After electrotransfer, wash the film with TBST at room temperature for 5 minutes;
    2. Incubate the film in the blocking solution for 1 hour at room temperature;
    3. Wash the film with TBST for 3 times, 5 minutes each time.
     
    Antibody incubation
    1. Use 5% skim milk powder to prepare the primary antibody working liquid (recommended dilution ratio for primary antibody 1:1000), gently shake and incubate with the film at 4°C overnight;
    2. Wash the film with TBST 3 times, 5 minutes each time;
    3. Add the secondary antibody to the blocking solution and incubate with the film gently at room temperature for 1 hour;
    4. After incubation, wash the film with TBST 3 times for 5 minutes each time.
     
    Antibody staining
    1. Add the prepared ECL luminescent substrate (or select other color developing substrate according to the second antibody) and mix evenly;
    2. Incubate with the film for 1 minute, remove excess substrate (keep the film moist), wrap with plastic film, and expose in the imaging system.
    IHC
    Experimental Protocol：
     
    Deparaffinization/Rehydration
    1. Deparaffinize/hydrate sections:
    2. Incubate sections in three washes of xylene for 5 min each.
    3. Incubate sections in two washes of 100% ethanol for 10 min each.
    4. Incubate sections in two washes of 95% ethanol for 10 min each.
    5. Wash sections two times in dH2O for 5 min each.
    6.Antigen retrieval: For Citrate: Heat slides in a microwave submersed in 1X citrate unmasking solution until boiling is initiated; continue with 10 min at a sub-boiling temperature (95°-98°C). Cool slides on bench top for 30 min.
     
    Staining
    1. Wash sections in dH2O three times for 5 min each.
    2. Incubate sections in 3% hydrogen peroxide for 10 min.
    3. Wash sections in dH2O two times for 5 min each.
    4. Wash sections in wash buffer for 5 min.
    5. Block each section with 100–400 µl of blocking solution for 1 hr at room temperature.
    6. Remove blocking solution and add 100–400 µl primary antibody diluent in to each section. Incubate overnight at 4°C.
    7. Remove antibody solution and wash sections with wash buffer three times for 5 min each.
    8. Cover section with 1–3 drops HRPas needed. Incubate in a humidified chamber for 30 min at room temperature.
    9. Wash sections three times with wash buffer for 5 min each.
    10. Add DAB Chromogen Concentrate to DAB Diluent and mix well before use.
    11. Apply 100–400 µl DAB to each section and monitor closely. 1–10 min generally provides an acceptable staining intensity.
    12. Immerse slides in dH2O.
    13. If desired, counterstain sections with hematoxylin.
    14. Wash sections in dH2O two times for 5 min each.
    15. Dehydrate sections: Incubate sections in 95% ethanol two times for 10 sec each; Repeat in 100% ethanol, incubating sections two times for 10 sec each; Repeat in xylene, incubating sections two times for 10 sec each.
    16. Mount sections with coverslips and mounting medium.
     

    Datasheet & SDS

    Biological Description

    Specificity
    Presenilin 1/PS-1 Rabbit mAb detects endogenous levels of total Presenilin 1/PS-1 protein.
    Subcellular Location
    Cell membrane, Cell projection, Endoplasmic reticulum, Endosome, Golgi apparatus, Membrane, Synapse
    Uniprot ID
    P49768
    Clone
    G24D17
    Synonym(s)
    AD3, PS1, PSNL1, PSEN1, Presenilin-1, PS-1, Protein S182
    Background
    Presenilin 1 (PSEN1) is a transmembrane protein belonging to the presenilin family and is composed of 467 amino acids, sharing approximately 65% sequence similarity with presenilin 2 (PSEN2). Structurally, PSEN1 contains nine transmembrane domains interconnected by hydrophilic loops that extend into either the extracellular space or the cytosol. It is a core component of the γ-secretase complex, which is essential for amyloid precursor protein (APP) processing and amyloid-β peptide generation. Within PSEN1, two conserved aspartic acid residues—Asp275 and Asp385, located in transmembrane domains 6 and 7—serve as catalytic sites required for both PSEN1 endoproteolysis and γ-secretase enzymatic function. The N-terminal region (TM1, HL1, and TM2) also plays a critical role in substrate catalysis within the γ-secretase complex. Beyond APP metabolism, PSEN1 regulates additional signaling pathways. Together with PSEN2, it modulates Notch signaling by mediating the intramembrane cleavage of Notch receptors. This cleavage event releases the Notch intracellular domain (NICD), which subsequently translocates into the nucleus to regulate cell fate decisions, differentiation, and proliferation. PSEN1 has also been implicated in Wnt signaling, where it influences β-catenin stability. Specifically, PSEN1 promotes β-catenin phosphorylation and inhibits the activity of cell cycle regulators such as cyclin D1, CDK6, and c-Myc. Mutations in PSEN1 can disrupt Wnt signaling in different ways—either suppressing the pathway, which contributes to amyloid-β–induced neurotoxicity and neuronal death, or aberrantly enhancing it, leading to abnormal cell-cycle re-entry and neurodegeneration. In addition, PSEN1 participates in the regulation of epidermal growth factor receptor (EGFR) trafficking and degradation. Loss of PSEN1 function has been associated with impaired EGFR transport from endosomes to lysosomes, thereby reducing receptor turnover. PSEN1 may also influence intracellular signaling by modulating phospholipase C (PLC) and protein kinase C (PKC) activity. Collectively, PSEN1 acts as a multifunctional regulator that integrates diverse molecular pathways, including APP processing, Notch signaling, Wnt signaling, EGFR trafficking, and kinase-mediated signaling cascades.
    References
    • https://pubmed.ncbi.nlm.nih.gov/36142879/

    Tech Support

    Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

    Handling Instructions

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