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TNFSF9 anticorps (Tumor Necrosis Factor (Ligand) Superfamily, Member 9) (AA 200-250) Primary Antibody

TNFSF9 Reactivité: Humain IF (p), IHC (p), WB Hôte: Lapin Polyclonal
Independent Validation (1)
N° du produit ABIN705056
$339.90
Plus shipping costs $45.00
100 μL
local_shipping Destination: Etats-Unis
Envoi sous 4 à 6 jours ouvrables
  • Antigène
    Épitope
    AA 200-250
    Reactivité
    Humain
    Hôte
    Lapin
    Clonalité
    Polyclonal
    Conjugué
    Cet anticorp TNFSF9 est non-conjugé
    Application
    Immunofluorescence (Paraffin-embedded Sections) (IF (p)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Western Blotting (WB)
    Purification
    Purified by Protein A.
    Immunogène
    KLH conjugated synthetic peptide derived from human TNFSF9
    Isotype
    IgG
  • Indications d'application
    WB 1:100-1000
    IHC-P 1:100-500
    IF(IHC-P) 1:50-200
    Restrictions
    For Research Use only
  • Validation #029578 (Immunohistochemistry)
    'Independent Validation' signe
    by
    Immunohistochemistry Core, NYU Langone
    No.
    #029578
    Date
    18.01.2014
    Antigène
    Numéro du lot
    999892W
    Application validée
    Immunohistochemistry
    Contrôle positif
    Human liver
    Contrôle négative
    Human placental decidual cells
    Conclusion
    Signal was detected in positive control tissue and not in negative control tissue.
    'Independent Validation' signe
    Validation Images
    Protocole
    Anticorps primaire
    • Antibody: Tumor necrosis factor ligand superfamily member 9 (TNFSF9)
    • Catalog number: ABIN705056
    • Lot number: 999892W
    Anticorps secondaire
    • Antibody: Biotinylated goat anti-rabbit/anti-mouse (Kit)
    • Lot number: D05923BA
    Full Protocol
    • Immunohistochemistry was performed on a Ventana NexES automated platform, instrument manufacturer specific reagents are italicized.
    • 1. Slides were preheated in convection oven at 60°C for 30 minutes
    • 2. Deparaffinization procedure: - 3 changes of Xylene, 5 minutes each - 3 changes of 100% Ethanol, 3 minutes each - 3 changes of 95% Ethanol, 3 minutes each - Rinsed in distilled water, 3 changes
    • 1. Heat retrieval procedure - Slides retrieved in 10.0 mM Citrate, pH6.0 in a 1000W microwave oven (~100°C) for 15 minutes. - Slides were allowed to cool (in citrate) for 30 minutes. - Slides were washed x 3 in Distilled water
    • 1. NexES instrument procedure, iVIEW DAB paraffin protocol (*abridged*): - Slide chamber warmed to 37°C
    • 1. Slides rinsed with *reaction buffer* x 3
    • 1. *iVIEW Inhibitor (H2O2)* applied and incubated for 4 minutes
    • 1. Slides rinsed with *reaction buffer*
    • 1. Antibody Application - Primary antibody diluted 1:250 in PBS (100 microliters applied/slide) - Ventana Isotype control applied neat - Slides incubated overnight at room temperature (~12 hours ~25°C)
    • 1. Slides rinsed with *reaction buffer* x3
    • 1. *iVIEW Biotinylated IgG* applied and incubated for 8 minutes
    • 1. Slides rinsed with *reaction buffer*
    • 1. *iVIEW Streptavidin-Horseradish Peroxidase* applied and incubated for 8 minutes
    • 1. Slides rinsed with *reaction buffer*
    • 1. *iVIEW DAB/H2O2* applied and incubated for 8 minutes
    • 1. Slides rinsed with *reaction buffer*
    • 1. *iVIEW Copper* applied and incubated for 4 minutes
    • 1. Slides rinsed with *reaction buffer*
    • 1. Slides washed in Dawn Detergent/tap water
    • 1. Counterstain Procedure - Hematoxylin (Leica 560 MX) 30 seconds - Slides washed in tap water, 1 minute - Decolorized (10% Acetic Acid in 70% ethanol), 1 minute - Slides washed in tap water, 1 minute - Bluing (Austin Clear Ammonia), 1 minute - Slides washed in tap water, 1 minute
    • 1. Dehydration/coverslipping procedure: - 3 changes of 95% Ethanol, 3 minutes each - 3 changes of 100% Ethanol, 3 minutes each - 3 changes of Xylene, 5 minutes each - Mounted with Permount
    • 1. Imaging: Leica SCN 400F Whole Slide Scanner with Digital Image Hub and Leica Slidepath software
    Notes
    • Deviations from protocol/procedure supplied by manufacturer:
    • Step 1: Heated tissue 60°C for 30 minutes; manufacturer heats for 45 minutes.
    • Step 2: No ethanol wash was performed during deparaffinization; manufacturer includes 1 wash of 80% ethanol for 3 minutes.
    • Step 3.1: Slides heated for 15 minutes; manufacturer provides a range of 15-20 minutes.
    • Step 3.2: Slides cooled for 30 minutes; manufacturer cools for 20 minutes.
    • Step 4: Italicized reagents and incubation time are fixed instrument parameters.
    • Step 5: Secondary species-specific serum block not used; manufacturer blocks with 5% normal goat serum for 2 hours.
    • Step 8.1: Antibody diluted in PBS at 1:250; manufacture recommends dilution range (1:100-1:500). No primary antibody diluent recommended.
    • Step 8.2.1: Primary antibody incubated at room temperature overnight; manufacturer incubates overnight 4°C with agitation.
    • Tissue Interpretation (limited): • TNFSF9: Under the staining parameters described above, placenta was weakly cytoplasmic positive (trophoblastic and decidua cells). Liver hepatocytes were weakly stained. Substantial signal detected in limited number of other tissues, including kidney, prostate, and skin.
    • • I-NC (Isotype negative control): No signal detected
    • • B-NC (Blank negative control): No signal detected
    • Signal Localization: - Signal to noise was adequate with predominately cytoplasmic subcellular localization observed. Nuclear staining was rare and no distinct membrane signal was detected.
  • Format
    Liquid
    Concentration
    1 μg/μL
    Buffer
    Aqueous buffered solution containing 1 % BSA, 50 % glycerol and 0.09 % sodium azide.
    Agent conservateur
    Sodium azide
    Précaution d'utilisation
    This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE, which should be handled by trained staff only.
    Stock
    -20 °C
    Stockage commentaire
    Store at -20°C
    Date de péremption
    12 months
  • Antigène
    Autre désignation
    Tnfsf9/Cd137l (TNFSF9 Antibody Extrait)
    Sujet

    Cytokine that binds to TNFRSF9. Induces the proliferation of activated peripheral blood T-cells. May have a role in activation-induced cell death (AICD). May play a role in cognate interactions between T-cells and B-cells/macrophages.

    Subcellular location: Extracellular

    Synonyms: CD137L, 4-1BB-L, Tumor necrosis factor ligand superfamily member 9, 4-1BB ligand, 4-1BBL, TNFSF9

    ID gène
    8744
    UniProt
    P41273
    Pathways
    Activated T Cell Proliferation, Cancer Immune Checkpoints
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