Rabbit Prohibitin/PHB Polyclonal Antibody | anti-PHB antibody

Anti-Prohibitin/PHB Antibody

Cat. Num. AAA19234

• Gene Names: PHB; PHB1
• Reactivity: Human, Mouse, Rat
• Applications: Western Blot, Immunohistochemistry, Flow Cytometry, Functional Assay, ELISA
• Purity: Immunogen affinity purified.
• Synonyms: Prohibitin/PHB, Antibody; Anti-Prohibitin/PHB Antibody; PHB; Prohibitin; prohibitin; anti-PHB antibody

• Host: Rabbit
• Reactivity: Human, Mouse, Rat
• Clonality: Polyclonal
• Isotype: Rabbit IgG
• Specificity: Rabbit IgG polyclonal antibody for Prohibitin/PHB detection.
• Purity/Purification: Immunogen affinity purified.
• Form/Format: Lyophilized. Each vial contains 4mg Trehalose, 0.9mg NaCl and 0.2mg Na2HPO4.

• Applicable Applications for anti-PHB antibody: Western Blot (WB), Immunohistochemistry-Paraffin (IHC-P), Flow Cytometry (FC/FACS/FCM), Direct ELISA (EIA)
• Application Notes: WB: 0.25-0.5ug/ml|Human, Mouse, Rat|; IHC-P: 2-5ug/ml|Human, Mouse, Rat|; FC/FACS/FCM: 1-3ug/1x106 cells|Rat|; Direct ELISA: 0.1-0.5ug/ml|Human|
• Immunogen: E Coli-derived human Prohibitin/PHB recombinant protein (Position: M1-I257).
• Reconstitution: Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
• Recommended Detection Systems: Recommended Detection Systems
• Preparation and Storage: Store at -20 degree C for one year. After reconstitution, at 4 degree C for one month. It can also be aliquotted and stored frozen at -20 degree C for a longer time. Avoid repeated freezing and thawing.

FCM (Flow Cytometry)

(Figure 10. Flow Cytometry analysis of RH35 cells using anti-Prohibitin/PHB antibody (AAA19234).Overlay histogram showing RH35 cells stained with AAA19234 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Prohibitin/PHB Antibody (AAA19234, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

IHC (Immunohistchemistry)

(Figure 9. IHC analysis of Prohibitin/PHB using anti-Prohibitin/PHB antibody (AAA19234).Prohibitin/PHB was detected in paraffin-embedded section of human rectal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-Prohibitin/PHB Antibody (AAA19234) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # with DAB as the chromogen.)

IHC (Immunohistochemistry)

(Figure 8. IHC analysis of Prohibitin/PHB using anti-Prohibitin/PHB antibody (AAA19234).Prohibitin/PHB was detected in paraffin-embedded section of human ovarian cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-Prohibitin/PHB Antibody (AAA19234) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # with DAB as the chromogen.)

IHC (Immunohistochemistry)

(Figure 7. IHC analysis of Prohibitin/PHB using anti-Prohibitin/PHB antibody (AAA19234).Prohibitin/PHB was detected in paraffin-embedded section of human liver cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-Prohibitin/PHB Antibody (AAA19234) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # with DAB as the chromogen.)

IHC (Immunohistchemistry)

(Figure 6. IHC analysis of Prohibitin/PHB using anti-Prohibitin/PHB antibody (AAA19234).Prohibitin/PHB was detected in paraffin-embedded section of human breast cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-Prohibitin/PHB Antibody (AAA19234) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # with DAB as the chromogen.)

IHC (Immunohistochemistry)

(Figure 5. IHC analysis of Prohibitin/PHB using anti-Prohibitin/PHB antibody (AAA19234).Prohibitin/PHB was detected in paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-Prohibitin/PHB Antibody (AAA19234) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # with DAB as the chromogen.)

IHC (Immunohistochemistry)

(Figure 4. IHC analysis of Prohibitin/PHB using anti-Prohibitin/PHB antibody (AAA19234).Prohibitin/PHB was detected in paraffin-embedded section of human adrenocortical adenoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-Prohibitin/PHB Antibody (AAA19234) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # with DAB as the chromogen.)

IHC (Immunohistochemistry)

(Figure 3. IHC analysis of Prohibitin/PHB using anti-Prohibitin/PHB antibody (AAA19234).Prohibitin/PHB was detected in paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-Prohibitin/PHB Antibody (AAA19234) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # with DAB as the chromogen.)

IHC (Immunohistochemistry)

(Figure 2. IHC analysis of Prohibitin/PHB using anti-Prohibitin/PHB antibody (AAA19234).Prohibitin/PHB was detected in paraffin-embedded section of mouse brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-Prohibitin/PHB Antibody (AAA19234) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # with DAB as the chromogen.)

WB (Western Blot)

(Figure 1. Western blot analysis of Prohibitin/PHB using anti-Prohibitin/PHB antibody (AAA19234).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30ug of sample under reducing conditions.Lane 1: human Hela whole cell lysatesLane 2: human Jurkat whole cell lysatesLane 3: human Hek293 whole cell lysatesLane 4: human K562 whole cell lysatesLane 5: human HT1080 whole cell lysatesLane 6: human Sw620 whole cell lysatesLane 7: human U87 whole cell lysatesLane 8: human A549 whole cell lysatesLane 9: rat brain tissue lysatesLane 10: rat heart tissue lysatesLane 11: rat liver tissue lysatesLane 12: rat PC-12 whole cell lysatesLane 13: mouse brain tissue lysatesLane 14: mouse heart tissue lysatesLane 15: mouse liver tissue lysatesLane 16: mouse NIH/3T3 whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with rabbit anti-Prohibitin/PHB antigen affinity purified polyclonal antibody (Catalog # AAA19234) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # with Tanon 5200 system. A specific band was detected for Prohibitin/PHB at approximately 28-30KD. The expected band size for Prohibitin/PHB is at 28-30KD.)

Related Product Information for anti-PHB antibody

PHB(Prohibitin), also known as PHB1, is a protein that in humans is encoded by the PHB gene. White et al. (1991) mapped the PHB gene to chromosome 17 by analysis of human-mouse somatic cell hybrid cell lines using a genomic fragment of human prohibitin DNA isolated from a library using the rat prohibitin cDNA clone. By in situ hybridization, they localized the gene to 17q21. Sato et al. (1992) isolated the human homolog of the rat prohibitin gene and mapped it to 17q12-q21 by in situ hybridization. Proliferation of tumor cells depends on new blood vessel formation(angiogenesis) that accompanies malignant progression. Anticancer therapies using angiogenesis inhibitors or cytotoxic agents targeted to the vasculature of tumors have been evaluated in clinical trials. Although white fat is a nonmalignant tissue, it has the capability to quickly proliferate and expand. Furthermore, it is highly vascularized. Rupnick et al. (2002) showed that nonspecific angiogenesis inhibitors can prevent the development of obesity of mice.

References

1. Rupnick, M. A., Panigrahy, D., Zhang, C. -Y., Dallabrida, S. M., Lowell, B. B., Langer, R., Folkman, M. J. Adipose tissue mass can be regulated through the vasculature. Proc. Nat. Acad. Sci. 99: 10730-10735, 2002.
2. Sato, T., Saito, H., Swensen, J., Olifant, A., Wood, C., Danner, D., Sakamoto, T., Takita, K., Kasumi, F., Miki, Y., Skolnick, M., Nakamura, Y. The human prohibitin gene located on chromosome 17q21 is mutated in sporadic breast cancer. Cancer Res. 52: 1643-1646, 1992.
3. White, J. J., Ledbetter, D. H., Eddy, R. L., Jr., Shows, T. B., Stewart, D. A., Nuell, M. J., Friedman, V., Wood, C. M., Owens, G. A., McClung, J. K., Danner, D. B., Morton, C. C. Assignment of the human prohibitin gene (PHB) to chromosome 17 and identification of a DNA polymorphism. Genomics 11: 228-230, 1991. Note: Erratum: Genomics 11: 732 only, 1991.

NCBI and Uniprot Product Information

• NCBI GI #: 246483
• NCBI GeneID: 5245
• UniProt Accession #: P35232; Q4VBQ0
• Molecular Weight: 29,804 Da
• NCBI Official Full Name: prohibitin
• NCBI Official Synonym Full Names: prohibitin
• NCBI Official Symbol: PHB
• NCBI Official Synonym Symbols: PHB1
• NCBI Protein Information: prohibitin
• UniProt Protein Name: Prohibitin
• UniProt Gene Name: PHB
• UniProt Entry Name: PHB_HUMAN