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Choisissez des Panels configurables & des Kits préconfigurés - OU - des MAPmate™ de signalisation cellulaire
Concevez vos kits MILLIPLEX® MAP et obtenez leur prix.
Panels configurables & Kits préconfigurés
Notre large gamme est constituée de panels multiplex qui vous permettent de choisir, au sein d'un panel, les analytes qui répondent le mieux à vos besoins. Sur un autre onglet, vous pouvez choisir un format cytokine préconfiguré ou un kit Simplex.
Kits de signalisation cellulaire & MAPmate™
Choisissez des kits préconfigurés qui permettent d'explorer l'ensemble des voies ou des processus. Ou concevez vos propres kits en choisissant des Simplex MAPmate™ et en suivant les instructions fournies.
Les MAPmate™ suivants ne peuvent pas être utilisés ensemble : -des MAPmate™ qui nécessitent des tampons différents -des paires de MAPmate™ totaux et phospho-spécifiques, par ex. GSK3β total et GSK3β (Ser 9) -des MAPmate™ PanTyr et spécifiques d'un site, par ex. Récepteur Phospho-EGF et phospho-STAT1 (Tyr701) -Plus d'un phospho-MAPmate™ pour une seule cible (Akt, STAT3). -GAPDH et β-Tubuline ne peuvent pas être utilisés avec les kits ou les MAPmate™ contenant panTyr.
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Sélectionner une espèce, un type de panel, un kit ou un type d'échantillon
Pour commencer à concevoir votre kit MILLIPLEX® MAP, sélectionnez une espèce, un type de panel ou un kit d'intérêt.
Custom Premix Selecting "Custom Premix" option means that all of the beads you have chosen will be premixed in manufacturing before the kit is sent to you.
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List
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96-Well Plate
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Ajouter des réactifs supplémentaires (Un kit "Buffer and Detection Kit" est nécessaire pour une utilisation avec les MAPmate™)
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48-602MAG
Buffer Detection Kit for Magnetic Beads
1 Kit
Option de gain de place Nos clients qui commandent plusieurs kits peuvent choisir d'économiser de l'espace de stockage en éliminant l'emballage de chaque kit et de recevoir les composants de leur essai multiplex conditionnés sous poches en plastique pour un stockage plus compact.
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PANSORBIN® Cells, Lyophilized : FDS (Fiches de données de sécurité), certificats d’analyse (CoA) et de qualité (CoQ), dossiers, brochures et autres documents disponibles.
Note that this data sheet is not lot-specific and is representative of the current specifications for this product. Please consult the vial label and the certificate of analysis for information on specific lots. Also note that shipping conditions may differ from storage conditions.
Revision
01-June-2009 JSW
Description
Formalin-fixed and heat-killed Staphylococcus aureus cells bearing protein A that bind to the Fc portion of human IgG subclasses 1, 2, and 4, and to IgG from other species (binding capacity: ≥2 mg human IgG/100 mg cells). Used for many purposes, including radioimmunoassays, enzyme immunoassays, agglutination, and immunoprecipitation of antigens. PANSORBIN® also stimulates B-lymphocyte production and inhibits neoplasm growth. Lyophilized from preparations of Cat. No. 507858. Note: size of 5 g or 1 g refers to the wet cell weight.
Form
Lyophilized
Formulation
Lyophilized from PBS, 0.1% NaN₃.
Intert gas (Yes/No)
Packaged under inert gas
Recommended reaction conditions
The procedures used for performing immunoprecipitations using PANSORBIN® cells are somewhat variable, but this protocol can serve as a general guideline.
PANSORBIN® cells work best when the antibody is human (IgG1, IgG2, IgG4), rabbit IgG (all isotypes), or mouse (IgG2a, IgG2b, IgG3).
Prewashing:
1. Transfer desired amount of PANSORBIN® cells to a microfuge tube (typically 10-30 µl of a 10% PANSORBIN® cell suspension per immunoprecipitation).
2. Centrifuge at 3000 x g for 5 min, or at maximum speed in a microfuge for 1 min at 4°C.
3. Aspirate the supernatant.
4. Resuspend to the original volume or in 1.0 ml of cold lysis buffer, whichever is greater. Mix gently.
5. Repeat wash (steps 2-4).
6. Resuspend the final pellet to the original volume (from step 1) in lysis buffer.
7. If kept sterile, washed PANSORBIN® cells are stable at 4°C.
Pre-Clearing (Optional):
Pre-clearing with PANSORBIN® cells prior to immunoprecipitation removes material that binds nonspecifically, reducing the background.
1. Add 10-30 µl of washed PANSORBIN® cells to the lysate prior to incubation with the primary antibody.
2. Incubate for 1 h on ice, mixing occasionally.
3. Centrifuge at 3000 x g for 5 min, or at maximum speed in a microfuge for 1 min at 4°C.
4. Transfer the supernatant to another microfuge tube and discard the pellet. Alternatively, the lysate can be pre-cleared with PANSORBIN® cells which have been pre-bound to nonspecific antiserum (see Secondary Antibody pre-binding procedure).
Primary Antibody:
1. Add the required amount of primary antibody to the lysate.
2. Incubate for at least 1 h on ice, mixing occasionally.
3. If a secondary antibody step is not required or desired, proceed to the PANSORBIN® Step described below.
Secondary Antibody:
In some cases, a secondary antibody will be necessary. For example, if the primary antibody is mouse IgG1, it is best to use a rabbit anti-mouse IgG as a secondary antibody. If desired, the secondary antibody can be pre-bound to the PANSORBIN® cells to eliminate the loss of antibody-antigen complexes which have not bound to the protein A.
1. Incubate the desired amount of secondary antibody with prewashed PANSORBIN® cells for 45-60 min on ice.
2. Centrifuge at 3000 x g for 5 min, or at maximum speed in a microfuge for 1 min at 4°C.
3. Aspirate the supernatant and wash the lysis buffer.
4. Resuspend in lysis buffer to a volume equal to the original volume of PANSORBIN® cells which were added.
Alternatively, the unbound secondary antibody and PANSORBIN® cells can be incubated with the sample sequentially, following primary antibody incubation.
PANSORBIN® Step:
1. Add at least 10X the amount of PANSORBIN® cells (alone or bound to secondary antibody) needed to precipitate the primary (or secondary) antibody. PANSORBIN® cells bind about 2 mg of human IgG/ml. Typically, add 10-30 µl of PANSORBIN® cells per immunoprecipitation.
This quantity gives a visible pellet.
2. Incubate for 1-2 h on ice, mixing regularly.
3. Centrifuge at 3000 x g for 5 min, or at maximum speed in a microfuge for 1 min at 4°C.
4. Wash PANSORBIN® pellet 2-3 times with lysis buffer.
5. If samples are to be loaded onto a polyacrylamide gel, the PANSORBIN® pellet can be resuspended in sample buffer, boiled for 10 min, centrifuged quickly (30 s in a microfuge at 20°C). Load the supernatant onto the gel. If a high background is observed, it may be necessary to pre-clear lysates, prebind the antibody to PANSORBIN® cells, or use a stronger lysis buffer (e.g. RIPA). This protocol is meant to serve as a guideline and may need to be modified for specific applications.
Solubility
Reconstitution with 10 ml sterile dH₂O yields a 10% (w/v) cell suspension.
Storage
+2°C to +8°C
Do Not Freeze
Ok to freeze
Special Instructions
Following reconstitution, aliquot and refrigerate (4°C). Stock solutions are stable for up to 1 month at 4°C.
Toxicity
Standard Handling
References
Kierszenbaum, F., et al. 1991. Immunology74, 317. Meikle, P.J., et al. 1991. J. Biol. Chem.266, 22569. Ezaki, O., et al. 1989. Biochem. Biophys. Res. Commun.159, 1368. Murakami, H., et al. 1988. Biochem. J.256, 917. Kessler, S.W. 1975. J. Immunol.115, 1617.
