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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.)

Rabbit Prohibitin/PHB Polyclonal Antibody | anti-PHB antibody

Anti-Prohibitin/PHB Antibody

Gene Names
PHB; PHB1
Reactivity
Human, Mouse, Rat
Applications
Western Blot, Immunohistochemistry, Flow Cytometry, Functional Assay, ELISA
Purity
Immunogen affinity purified.
Synonyms
Prohibitin/PHB; Polyclonal Antibody; Anti-Prohibitin/PHB Antibody; PHB; Prohibitin; prohibitin; anti-PHB antibody
Ordering
For Research Use Only!
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.)

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 (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 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 (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 (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 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 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 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.)

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.)

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 #
NCBI GeneID
UniProt Accession #
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

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Product Notes

The PHB phb (Catalog #AAA19234) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-Prohibitin/PHB Antibody reacts with Human, Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's Prohibitin/PHB can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunohistochemistry-Paraffin (IHC-P), Flow Cytometry (FC/FACS/FCM), Direct ELISA (EIA). 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|. Researchers should empirically determine the suitability of the PHB phb for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "Prohibitin/PHB, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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