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PC66 Anti-Bax (Ab-1) (150-165) Rabbit pAb

PC66
  
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      Overview

      Replacement Information

      Key Spec Table

      Host
      Rb
      Description
      Overview

      This product has been discontinued.



      Recognizes the ~21 kDa monomeric and ~44 -50 kDa dimeric Bax protein in induced HeLa and MCF-7 cell lysates.
      Catalogue NumberPC66
      Brand Family Calbiochem®
      References
      ReferencesBargou, R.C., et al. 1995. Int. J. Cancer 60, 854.
      Hanada, M., et al. 1995. J. Biol. Chem. 270, 11962.
      Miyashita, T. and Reed, J.C. 1995. Cell 80, 293.
      Korsmeyer, S.J., et al. 1993. Semin. Cancer Biol. 4, 327.
      Oltvai, Z.N., et al. 1993. Cell 74, 609.
      Product Information
      FormLiquid
      FormulationIn 0.05 M sodium phosphate buffer, 0.2% gelatin.
      Negative controlUninduced HeLa or MCF-7 cells
      Positive controlDoxorubicin-treated HeLa or MCF7 cells, mouse intestinal tissue, or human lymph node tissue
      Preservative≤0.1% sodium azide
      Quality LevelMQ100
      Applications
      Key Applications Frozen Sections
      Immunoblotting (Western Blotting)
      Paraffin Sections
      Application NotesFrozen Sections (2-5 µg/ml)
      Immunoblotting (0.5-5 µg/ml)
      Paraffin Sections (2-5 µg/ml, pressure cooker pre-treatment required)
      Application CommentsDoes 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
      Immunogena synthetic peptide (GWIQDQGGWDGLLSYF) corresponding to amino acids 150-165 of human Bax
      ImmunogenHuman
      HostRabbit
      IsotypeIgG
      Concentration Label Please refer to vial label for lot-specific concentration
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Storage and Shipping Information
      Ship Code Blue Ice Only
      Toxicity Standard Handling
      Storage +2°C to +8°C
      Do not freeze Yes
      Packaging Information
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      Anti-Bax (Ab-1) (150-165) Rabbit pAb Certificates of Analysis

      TitleLot Number
      PC66

      References

      Reference overview
      Bargou, R.C., et al. 1995. Int. J. Cancer 60, 854.
      Hanada, M., et al. 1995. J. Biol. Chem. 270, 11962.
      Miyashita, T. and Reed, J.C. 1995. Cell 80, 293.
      Korsmeyer, S.J., et al. 1993. Semin. Cancer Biol. 4, 327.
      Oltvai, Z.N., et al. 1993. Cell 74, 609.

      Brochure

      Title
      Caspases and other Apoptosis Related Tools Brochure
      Data Sheet

      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.

      Revision15-October-2007 RFH
      ApplicationFrozen Sections (2-5 µg/ml)
      Immunoblotting (0.5-5 µg/ml)
      Paraffin Sections (2-5 µg/ml, pressure cooker pre-treatment required)
      DescriptionPurified rabbit polyclonal antibody. Recognizes the ~21 kDa bax protein.
      BackgroundBax 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.
      HostRabbit
      Immunogen speciesHuman
      Immunogena synthetic peptide (GWIQDQGGWDGLLSYF) corresponding to amino acids 150-165 of human Bax
      IsotypeIgG
      Specieshuman, mouse, opossum, rat
      Positive controlDoxorubicin-treated HeLa or MCF7 cells, mouse intestinal tissue, or human lymph node tissue
      Negative controlUninduced HeLa or MCF-7 cells
      FormLiquid
      FormulationIn 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
      CommentsDoes 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
      ReferencesBargou, R.C., et al. 1995. Int. J. Cancer 60, 854.
      Hanada, M., et al. 1995. J. Biol. Chem. 270, 11962.
      Miyashita, T. and Reed, J.C. 1995. Cell 80, 293.
      Korsmeyer, S.J., et al. 1993. Semin. Cancer Biol. 4, 327.
      Oltvai, Z.N., et al. 1993. Cell 74, 609.