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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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48-602MAG
Buffer Detection Kit for Magnetic Beads
1 Kit
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Recognizes the ~21 kDa monomeric and ~44 -50 kDa dimeric Bax protein in induced HeLa and MCF-7 cell lysates.
Catalogue Number
PC66
Brand Family
Calbiochem®
References
References
Bargou, R.C., et al. 1995. Int. J. Cancer60, 854. Hanada, M., et al. 1995. J. Biol. Chem.270, 11962. Miyashita, T. and Reed, J.C. 1995. Cell80, 293. Korsmeyer, S.J., et al. 1993. Semin. Cancer Biol.4, 327. Oltvai, Z.N., et al. 1993. Cell74, 609.
Product Information
Form
Liquid
Formulation
In 0.05 M sodium phosphate buffer, 0.2% gelatin.
Negative control
Uninduced HeLa or MCF-7 cells
Positive control
Doxorubicin-treated HeLa or MCF7 cells, mouse intestinal tissue, or human lymph node tissue
Does not detect a bax-sized band in bax knockout mice and does not react to other members of the bcl-2 family. Antibody should be titrated for optimal results in individual systems.
Immunoblotting Use a 0.2 µm filter to minimize loss of bax through the membrane. Be sure to use a positive control with each blot. We usually detect extra high molecular weight bands but do not see other bands near bax (~22 kDa). The extra bands appear to be nonspecific interactions with unknown proteins. A band at ~44-50 kDa is common. The number and intensity of extra bands can be minimized by using 0.3% Tween®-20 detergent in the antibody and wash buffers.
Controls Bax can be induced using standard procedures. Treating with doxorubicin (adriamycin) for 48 h works well for HeLa, MCF-7 and H1299 cells. Untreated HL-60 and HS27 cells do not express bax and can be used as negative controls. Normal colon sections can be used as a positive control for immunohistochemistry. Alternatively, you can spot the immunogen peptide (Cat. No. PP51) on the membrane as a positive control. After the proteins have transferred, let the membrane dry and add approximately 0.1 µg peptide (in approximately 1 µl PBS) to a corner of the membrane (outside the area of gel protein transfer). As additional controls, spot 1 µl of primary antibody and 1 µl of secondary antibody. Let the spot(s) air dry, then proceed with the rest of the protocol. All spots should give a very strong positive signal. Note: the peptide Cat. No. PP51 is too small to be used for gel electrophoresis.
Biological Information
Immunogen
a synthetic peptide (GWIQDQGGWDGLLSYF) corresponding to amino acids 150-165 of human Bax
Immunogen
Human
Host
Rabbit
Isotype
IgG
Concentration Label
Please refer to vial label for lot-specific concentration
Bargou, R.C., et al. 1995. Int. J. Cancer60, 854. Hanada, M., et al. 1995. J. Biol. Chem.270, 11962. Miyashita, T. and Reed, J.C. 1995. Cell80, 293. Korsmeyer, S.J., et al. 1993. Semin. Cancer Biol.4, 327. Oltvai, Z.N., et al. 1993. Cell74, 609.
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.
Purified rabbit polyclonal antibody. Recognizes the ~21 kDa bax protein.
Background
Bax is a ~21 kDa protein with extensive amino acid homology with bcl-2. The protein is encoded by six exons and has been shown to undergo alternative splicing leading to at least two cytoplasmic forms. Bax has been shown to form heterodimers with bcl-2; the ratio of bcl 2/bax determines the survival or death of cells following an apoptotic stimulus such as removal of growth factor. Stimulation of bax synthesis also appears to be a result of wild type (but not mutant) p53 activity based on the observation that the bax gene promoter region contains 4 motifs showing consensus with p53 binding sites. bax binds downstream of the C domain of bcl-2 around amino acids 197-218 and the failure to do so results in onset of apoptosis, as does the formation of bax homodimers. Although the formation of the bcl-2/bax heterodimer appears to promote cell survival this may not be absolutely sufficient for the anti-cell death function. More recently it has been suggested that dysregulation of apoptosis due to imbalances in bax/bcl-2 levels may contribute to the pathogenesis of breast cancer.
Host
Rabbit
Immunogen species
Human
Immunogen
a synthetic peptide (GWIQDQGGWDGLLSYF) corresponding to amino acids 150-165 of human Bax
Isotype
IgG
Species
human, mouse, opossum, rat
Positive control
Doxorubicin-treated HeLa or MCF7 cells, mouse intestinal tissue, or human lymph node tissue
Negative control
Uninduced HeLa or MCF-7 cells
Form
Liquid
Formulation
In 0.05 M sodium phosphate buffer, 0.2% gelatin.
Concentration Label
Please refer to vial label for lot-specific concentration
Preservative
≤0.1% sodium azide
Comments
Does not detect a bax-sized band in bax knockout mice and does not react to other members of the bcl-2 family. Antibody should be titrated for optimal results in individual systems.
Immunoblotting Use a 0.2 µm filter to minimize loss of bax through the membrane. Be sure to use a positive control with each blot. We usually detect extra high molecular weight bands but do not see other bands near bax (~22 kDa). The extra bands appear to be nonspecific interactions with unknown proteins. A band at ~44-50 kDa is common. The number and intensity of extra bands can be minimized by using 0.3% Tween®-20 detergent in the antibody and wash buffers.
Controls Bax can be induced using standard procedures. Treating with doxorubicin (adriamycin) for 48 h works well for HeLa, MCF-7 and H1299 cells. Untreated HL-60 and HS27 cells do not express bax and can be used as negative controls. Normal colon sections can be used as a positive control for immunohistochemistry. Alternatively, you can spot the immunogen peptide (Cat. No. PP51) on the membrane as a positive control. After the proteins have transferred, let the membrane dry and add approximately 0.1 µg peptide (in approximately 1 µl PBS) to a corner of the membrane (outside the area of gel protein transfer). As additional controls, spot 1 µl of primary antibody and 1 µl of secondary antibody. Let the spot(s) air dry, then proceed with the rest of the protocol. All spots should give a very strong positive signal. Note: the peptide Cat. No. PP51 is too small to be used for gel electrophoresis.
Storage
+2°C to +8°C
Do Not Freeze
Yes
Toxicity
Standard Handling
References
Bargou, R.C., et al. 1995. Int. J. Cancer60, 854. Hanada, M., et al. 1995. J. Biol. Chem.270, 11962. Miyashita, T. and Reed, J.C. 1995. Cell80, 293. Korsmeyer, S.J., et al. 1993. Semin. Cancer Biol.4, 327. Oltvai, Z.N., et al. 1993. Cell74, 609.