Humain/Mouse MAPK Phosphorylation Array

N° du produit ABIN4955901

Détail du produit

Reactivité: Humain, Souris

Méthode de détection: Chemiluminescent

Type de méthode: Sandwich ELISA

Application: Antibody Array (AA)

Fonction: C-Series Human/Mouse MAPK Phosphorylation Antibody Array 1 Kit. Detects 17 Human/mouse proteins. Suitable for all liquid sample types but intended for use with cell and tissue lysates.

Type d'échantillon: Plasma, Cell Culture Supernatant, Serum, Cell Lysate, Tissue Lysate

Analytical Method: Semi-Quantitative

Specificité: Akt (P-Ser473), CREB (P-Ser133), ERK1 (P-T202/Y204)/ERK2 (P-Y185/Y187), GSK3a (P-Ser21), GSK3b (P-Ser9), HSP27 (P-Ser82), JNK (P-Thr183), MEK (P-Ser217/Ser221), MKK3 (P-Ser189), MKK6 (P-Ser207), MSK2 (P-Ser360), mTOR (P-Ser2448), p38 (P-Thr180/Tyr182), P53 (P-Ser15), P70S6K (P-Thr421/Ser424), RSK1 (P-Ser380), RSK2 (P-Ser386)

Attributs du produit:

• Easy to use
• No specialized equipment needed
• Compatible with nearly any liquid sample
• Proven technology (many publications)
• Highly sensitive (pg/mL)
• Sandwich ELISA specificity
• Higher density than ELISA, Western blot or bead-based multiplex

Ingrédients: Antibody Array Membranes
Biotinylated Detection Antibody Cocktail
Blocking Buffer
Wash Buffers 1 and 2
Cell & Tissue Lysis Buffer
Detection Buffers C and D
Plastic Incubation Tray
Protease Inhibitor Cocktail (in select kits)

Matériel non inclus: Pipettors, pipet tips and other common lab consumables
Orbital shaker or oscillating rocker
Tissue Paper, blotting paper or chromatography paper
Adhesive tape or Saran Wrap
Distilled or de-ionized water
A chemiluminescent blot documentation system (such as UVP's ChemiDoc-It® or EpiChem II Benchtop Darkroom), X-ray Film and a suitable film processor, or another chemiluminescent detection system.

Information d'application

Indications d'application: Perform ALL incubation and wash steps under gentle rotation or rocking motion (~0.5 to 1 cycle/sec) using an orbital shaker or oscillating rocker to ensure complete and even reagent/sample coverage. Rocking/rotating too vigorously may cause foaming or bubbles to appear on the membrane surface which, should be avoided. All washes and incubations should be performed in the Incubation Tray (ITEM 10) provided in the kit. Cover the Incubation Tray with the lid provided during all incubation steps to avoid evaporation and outside debris contamination. Ensure the membranes are completely covered with sufficient sample or reagent volume during each incubation. Avoid forceful pipetting directly onto the membrane, instead, gently pipette samples and reagents into a corner of each well. Aspirate samples and reagents completely after each step by suctioning off excess liquid with a pipette. Tilting the tray so the liquid moves to a corner and then pipetting is an effective method. Optional overnight incubations may be performed for the following step to increase overall spot signal intensities:
- Sample Incubation
- Biotinylated Antibody Cocktail Incubation
- HRP-Streptavidin Incubation

Commentaires:

The C-Series arrays feature chemiluminescent signal detection. The antibodies are spotted on nitrocellulose membrane solid supports and are handled in a very similar manner to Western blots.
All C-Series arrays work on the sandwich ELISA principle, utilizing a matched pair of antibodies: an immobilized capture antibody and a corresponding biotinylated detection antibody.

Volume d'échantillon: 1 mL

Plaque: Membrane

Protocole:

1. Block membranes
2. Incubate with Sample
3. Incubate with Biotinylated Detection Antibody Cocktail
4. Incubate with HRP-Conjugated Streptavidin
5. Incubate with Detection Buffers
6. Image with chemiluminescent imaging system
7. Perform densitometry and analysis

Restrictions: For Research Use only

Stockage

Conseil sur la manipulation: The antibody printed side of each membrane is marked by a dash (-) or number (#) in the upper left corner. Do not allow membranes to dry out during the experiment or they may become fragile and break OR high and/or uneven background may occur. Grasp membranes by the corners or edges only using forceps. DO NOT touch printed antibody spots.

Stock: -20 °C

Stockage commentaire: For best results, store the entire kit frozen at -20°C upon arrival. Stored frozen, the kit will be stable for at least 6 months which is the duration of the product warranty period. Once thawed, store array membranes and 1X Blocking Buffer at -20°C and all other reagents undiluted at 4°C for no more than 3 months.

Référence

Schuett, Schuett, Oberoi, Koch, Pretzer, Luchtefeld, Schieffer, Grote: "NADPH oxidase NOX2 mediates TLR2/6-dependent release of GM-CSF from endothelial cells." dans: FASEB journal : official publication of the Federation of American Societies for Experimental Biology, Vol. 31, Issue 6, pp. 2612-2624, (2017) (PubMed).