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Wählen Sie konfigurierbare Panels & Premixed-Kits - ODER - Kits für die zelluläre Signaltransduktion & MAPmates™
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Kits für die zelluläre Signaltransduktion & MAPmates™
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Die folgenden MAPmates™ sollten nicht zusammen analysiert werden: -MAPmates™, die einen unterschiedlichen Assaypuffer erfordern. -Phosphospezifische und MAPmate™ Gesamtkombinationen wie Gesamt-GSK3β und Gesamt-GSK3β (Ser 9). -PanTyr und locusspezifische MAPmates™, z.B. Phospho-EGF-Rezeptor und Phospho-STAT1 (Tyr701). -Mehr als 1 Phospho-MAPmate™ für ein einziges Target (Akt, STAT3). -GAPDH und β-Tubulin können nicht mit Kits oder MAPmates™, die panTyr enthalten, analysiert werden.
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Gewähltes Kit
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96-Well Plate
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Weitere Reagenzien hinzufügen (MAPmates erfordern die Verwendung eines Puffer- und Detektionskits)
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48-602MAG
Buffer Detection Kit for Magnetic Beads
1 Kit
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A human breast adenocarcinoma cell line with an epithelial cell morphology that was originally isolated from a pleural effusion. Useful as a positive control Western blotting of various proteins, such as estrogen receptors and the insulin-like growth factor binding proteins (IGFBP) BP-2, BP-4, and BP-5.
Laemmli Sample Buffer (SDS-PAGE) Resuspend the frozen cell pellet by adding 1.0 ml of Laemmli sample buffer (containing fresh 2-mercaptoethanol, 2-ME, at a final concentration of 500 mM) to the vial containing the cell pellet followed by gentle vortexing. (Alternatively dithiothreitol (DTT) to a final concentration of 100 mM may be substituted for the 2-ME). Heat the sample for 5 min at 90°C. Apply 15-25 µl to each lane of the gel. This is a starting amount, as actual amounts for optimal protein detection need to be experimentally determined for every antibody. Freeze the unused material in 25 µl aliquots and store at -70°C. (Before using the frozen samples, fresh 2-ME or DTT should be added and the sample heated for 5 min at 90°C).
Lane's Sample Buffer (IP/Immunoblot applications) Resuspend the frozen pellet by adding 1.0 ml of Lane's lysis buffer (150 mM NaCl, 50 mM Tris-HCl, pH 8.0, 5 mM EDTA, and 1% NP40) to the vial containing the cell pellet followed by gentle vortexing. Fresh 2-mercaptoethanol, 2-ME, must be added to the vial at a concentration of 500 mM prior to heating the sample for 5 min at 90°C. (Alternatively dithiothreitol (DTT) to a final concentration of 100 mM may be substituted for the 2-ME). Apply 15-25 µl to each lane of the gel. This is a starting amount, as actual amounts for optimal protein detection need to be experimentally determined for every antibody. Freeze the unused material in 25 µl aliquots and store at -70°C. (Before using the frozen samples, fresh 2-ME or DTT should be added and the sample heated for 5 min at 90°C).
Biological Information
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Storage and Shipping Information
Ship Code
Dry Ice Only
Toxicity
Standard Handling
Storage
≤ -70°C
Avoid freeze/thaw
Avoid freeze/thaw
Do not freeze
Ok to freeze
Special Instructions
Store at -70°C until lysed as described below. Following resuspension, store unused material in 25 µl aliquots at -70°C until needed. Do not expose to repeated cycles of freezing and thawing.
Packaging Information
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Bestellnummer
GTIN
WB39
0
Documentation
MCF7 Cell Pellet Analysenzertifikate
Titel
Chargennummer
WB39
Datenblatt
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
25-October-2007 RFH
Description
The Calbiochem® Cell Pellets are a collection of frozen cell pellets representing cell lines most commonly used for internal controls in immunoblotting and IP/immunoblotting applications. Cells are grown under standard conditions to a density of approximately 80-85% confluency, washed extensively to remove excess culture medium derived proteins (predominantly serum albumin) and then pelleted and flash frozen. Pellets are designed to be lysed by the addition of Laemmli sample buffer for SDS-PAGE applications. Alternatively, the pellet may be lysed using alternative buffers such as Lane's Buffer or specific buffers with lower concentrations of detergent for IP/immunoblotting applications. Vial containing 10 X 106 of the indicated cell type, flash frozen at -70°C.
Background
A human breast adenocarcinoma with an epithelial cell morphology that was originally isolated from a pleural effusion. This cell line expressed high levels of estrogen receptors and the insulin-like growth factor binding proteins (IGFBP) BP-2; BP-4; and BP-5. To preserve phosphorylation, orthovanadate should be included in the lysis buffer.
Form
Flash-frozen cell pellet
Comments
Suggested Preparation for Use
Laemmli Sample Buffer (SDS-PAGE) Resuspend the frozen cell pellet by adding 1.0 ml of Laemmli sample buffer (containing fresh 2-mercaptoethanol, 2-ME, at a final concentration of 500 mM) to the vial containing the cell pellet followed by gentle vortexing. (Alternatively dithiothreitol (DTT) to a final concentration of 100 mM may be substituted for the 2-ME). Heat the sample for 5 min at 90°C. Apply 15-25 µl to each lane of the gel. This is a starting amount, as actual amounts for optimal protein detection need to be experimentally determined for every antibody. Freeze the unused material in 25 µl aliquots and store at -70°C. (Before using the frozen samples, fresh 2-ME or DTT should be added and the sample heated for 5 min at 90°C).
Lane's Sample Buffer (IP/Immunoblot applications) Resuspend the frozen pellet by adding 1.0 ml of Lane's lysis buffer (150 mM NaCl, 50 mM Tris-HCl, pH 8.0, 5 mM EDTA, and 1% NP40) to the vial containing the cell pellet followed by gentle vortexing. Fresh 2-mercaptoethanol, 2-ME, must be added to the vial at a concentration of 500 mM prior to heating the sample for 5 min at 90°C. (Alternatively dithiothreitol (DTT) to a final concentration of 100 mM may be substituted for the 2-ME). Apply 15-25 µl to each lane of the gel. This is a starting amount, as actual amounts for optimal protein detection need to be experimentally determined for every antibody. Freeze the unused material in 25 µl aliquots and store at -70°C. (Before using the frozen samples, fresh 2-ME or DTT should be added and the sample heated for 5 min at 90°C).
Storage
Avoid freeze/thaw
≤ -70°C
Do Not Freeze
Ok to freeze
Special Instructions
Store at -70°C until lysed as described below. Following resuspension, store unused material in 25 µl aliquots at -70°C until needed. Do not expose to repeated cycles of freezing and thawing.