Anti-ATP-binding cassette sub-family A member 3 Mouse Antibody [F19C21]

Catalog No.: F2545

    Application: Reactivity:
    • Immunohistochemical analysis of formalin fixed paraffin embedded mouse lung tissue with F2545 at 1:100 dilution.

    Usage Information

    Dilution
    1:10
    Application
    IHC
    Reactivity
    Mouse, Rat
    Source
    Mouse
    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
    Positive Control Mouse lung; Rat lung
    Negative Control

    Exprimental Methods

    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
    ATP-binding cassette sub-family A member 3 Mouse mAb detects endogenous levels of total ATP-binding cassette sub-family A member 3 protein.
    Subcellular Location
    Cytoplasmic vesicle, Endosome, Lysosome, Membrane
    Uniprot ID
    Q99758
    Clone
    F19C21
    Synonym(s)
    Phospholipid-transporting ATPase ABCA3, ATP-binding cassette sub-family A member 3, Xenobiotic-transporting ATPase ABCA3, Abca3
    Background
    ATP-binding cassette sub-family A member 3 (ABCA3) is a highly conserved, multi-pass transmembrane glycoprotein in the ABC transporter superfamily that uses ATP hydrolysis to transport phospholipids into lamellar bodies (LBs) of alveolar type II cells, where pulmonary surfactant is assembled and stored. Structurally, ABCA3 consists of two similar halves, each containing six transmembrane domains and a cytosolic nucleotide-binding domain (NBD) with Walker A, Walker B, and ABCA-specific signature motifs, plus two essential N-linked glycosylation sites at Asn124 and Asn140 required for proper folding, stability, and LB targeting. Although expressed in multiple tissues, ABCA3 is highly enriched in the lung, where it is critical for LB biogenesis and surfactant homeostasis, particularly in transporting phosphatidylcholine, phosphatidylglycerol, and cholesterol. Mutations in ABCA3 can cause trafficking or functional defects, leading to neonatal surfactant deficiency, interstitial lung disease, and fatal respiratory distress.
    References
    • https://pubmed.ncbi.nlm.nih.gov/28025703/

    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

    Tel: +1-832-582-8158 Ext:3
    If you have any other enquiries, please leave a message.

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