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

Anti-Bax (Ab-1) (150-165) Rabbit pAb MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
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
Global Trade Item Number
Catalogue Number GTIN
PC66 0

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.