Product Details
Clone: | CB5.4 |
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Host: | Rat |
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Isotype: | IgG2a |
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Immunogen: | Recombinant mouse perforin (aa 98-534). |
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UniProt ID: | P10820 |
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Source: | Purified from ascites. |
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Species reactivity: | Mouse
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Specificity: | Recognizes an epitope in the region aa 402-534. |
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Applications: | ICC, IHC (FS), IHC (PS), IP, WB
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Formulation: | Liquid. In PBS containing 0.02% sodium azide. |
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Use/Stability: | Stable for at least 6 months after receipt when stored at +4°C. |
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Handling: | Avoid freeze/thaw cycles. |
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Shipping: | Blue Ice |
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Short Term Storage: | +4°C |
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Long Term Storage: | -20°C |
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Regulatory Status: | RUO - Research Use Only |
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Figure: Detection of perforin in the mouse T cell clone B6.1. (lane 1). Absence of perforin in the mouse fibroblast cell line NIH.3T3 (lane 2). The protein migrates as a 66 kDa species.
Method: Cell extracts from the T cell clone B6.1 (2x106) were resolved by SDS-PAGE under reducing conditions, transferred to nitrocellulose and probed with the CB5.4 antibody at 1 µg/ml. Proteins were visualized using a peroxidase-conjugated antibody to rat IgG and a chemiluminescence detection system.
Schematic structure of mouse perforin (recombinant, aa. 98-534). The antibody recognizes an epitope within aa. 402-534.
Figure: Detection of perforin in an individual mouse T cell. Perforin is located in cytoplasmic granules.
Method: A mouse T cell clone was plated onto polylysine treated glass slides, fixed and permeabilized in methanol at -20oC for 5 min, then in acetone at -20oC for 30 sec. After 3 washings in PBS, 0.1% BSA, slides were incubated with 5 µg/ml of CB5.4 antibody, washed in PBS, incubated in anti-rat IgG-HRP, washed in PBS, and developed in appropriate substrate solution (DAB or AEC).
Western Blot analysis of Perforin (mouse) mAb (CB5.4) (Prod. No. ALX-804-057): Lane 1: MW Marker, Lane 2: CTLL-2 cell lysate, Lane 3: 3T3 cell lysate. Western Blot analysis of Perforin 1 expression in CTLL-2 cell lysate and absence of perforin in the mouse fibroblast cell line 3T3.
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Product Literature References
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Th cells promote CTL survival and memory via acquired pMHC-I and endogenous IL-2 and CD40L signaling and by modulating apoptosis-controlling pathways: C.S. Umeshappa, et al.; PLoS One
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The Rac activator DOCK2 regulates natural killer cell-mediated cytotoxicity in mice through the lytic synapse formation: Y. Sakai, et al.; Blood
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Resistance of CTL to perforin-mediated lysis. Evidence for a lymphocyte membrane protein interacting with perforin: C. Muller & J. Tschopp; J. Immunol.
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The calcium-binding protein calreticulin is a major constituent of lytic granules in cytolytic T lymphocytes: M. Dupuis, et al.; J. Exp. Med.
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General Literature References
Fas-Fas ligand interactions are essential for the binding to and killing of activated macrophages by gamma delta T cells: J.E. Dalton, et al.; J. Immunol.
173, 3660 (2004),
Abstract;
Cubilin, a binding partner for galectin-3 in the murine utero-placental complex: S. Crider-Pirkle, et al.; J. Biol. Chem.
277, 15904 (2002),
Abstract;