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Product Info Summary
| SKU: | A01178-1 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IHC, WB |
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Product info
Product Name
Anti-Prohibitin/PHB Antibody Picoband®
View all Prohibitin Antibodies
SKU/Catalog Number
A01178-1
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Prohibitin/PHB Antibody Picoband® catalog # A01178-1. Tested in ELISA, Flow Cytometry, IHC, WB applications. This antibody reacts with Human, Mouse, Rat. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
Storage & Handling
Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
Cite This Product
Anti-Prohibitin/PHB Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A01178-1)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl and 0.2mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human Prohibitin/PHB recombinant protein (Position: M1-I257).
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A01178-1 is reactive to PHB in Human, Mouse, Rat
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Observed Molecular Weight
28-30 kDa
Calculated molecular weight
29.804kDa
Background of Prohibitin
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.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
A01178-1 is guaranteed for ELISA, Flow Cytometry, IHC, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.25-0.5μg/ml, Human, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human, Mouse, Rat
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human, Rat
ELISA, 0.1-0.5μg/ml, -
Positive Control
WB: human Hela whole cell, human Jurkat whole cell, human Hek293 whole cell, human K562 whole cell, human HT1080 whole cell, human Sw620 whole cell, human U87 whole cell, human A549 whole cell, rat brain tissue, rat heart tissue, rat liver tissue, rat PC-12 whole cell, mouse brain tissue, mouse heart tissue, mouse liver tissue, mouse NIH/3T3 whole cell
IHC: mouse brain tissue, rat brain tissue, human adrenocortical adenoma tissue, human lung cancer tissue, human breast cancer tissue, human liver cancer tissue, human ovarian cancer tissue, human rectal cancer tissue
FCM: RH35 cell, HL-60 cell
Validation Images & Assay Conditions
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Figure 1. Western blot analysis of Prohibitin/PHB using anti-Prohibitin/PHB antibody (A01178-1).
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 lysates,
Lane 2: human Jurkat whole cell lysates,
Lane 3: human Hek293 whole cell lysates,
Lane 4: human K562 whole cell lysates,
Lane 5: human HT1080 whole cell lysates,
Lane 6: human Sw620 whole cell lysates,
Lane 7: human U87 whole cell lysates,
Lane 8: human A549 whole cell lysates,
Lane 9: rat brain tissue lysates,
Lane 10: rat heart tissue lysates,
Lane 11: rat liver tissue lysates,
Lane 12: rat PC-12 whole cell lysates,
Lane 13: mouse brain tissue lysates,
Lane 14: mouse heart tissue lysates,
Lane 15: mouse liver tissue lysates,
Lane 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 # A01178-1) at 0.5 μg/mL overnight at 4°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 # EK1002) 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.
Click image to see more details
Figure 2. IHC analysis of Prohibitin/PHB using anti-Prohibitin/PHB antibody (A01178-1).
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 (A01178-1) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 3. IHC analysis of Prohibitin/PHB using anti-Prohibitin/PHB antibody (A01178-1).
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 (A01178-1) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 4. IHC analysis of Prohibitin/PHB using anti-Prohibitin/PHB antibody (A01178-1).
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 (A01178-1) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 5. IHC analysis of Prohibitin/PHB using anti-Prohibitin/PHB antibody (A01178-1).
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 (A01178-1) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 6. IHC analysis of Prohibitin/PHB using anti-Prohibitin/PHB antibody (A01178-1).
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 (A01178-1) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 7. IHC analysis of Prohibitin/PHB using anti-Prohibitin/PHB antibody (A01178-1).
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 (A01178-1) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 8. IHC analysis of Prohibitin/PHB using anti-Prohibitin/PHB antibody (A01178-1).
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 (A01178-1) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 9. IHC analysis of Prohibitin/PHB using anti-Prohibitin/PHB antibody (A01178-1).
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 (A01178-1) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 10. Flow Cytometry analysis of RH35 cells using anti-Prohibitin/PHB antibody (A01178-1).
Overlay histogram showing RH35 cells stained with A01178-1 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Prohibitin/PHB Antibody (A01178-1, 1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Click image to see more details
Figure 11. Flow Cytometry analysis of HL-60 cells using anti-Prohibitin/PHB antibody (A01178-1).
Overlay histogram showing HL-60 cells stained with A01178-1 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Prohibitin/PHB Antibody (A01178-1, 1 μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Protein Target Info & Infographic
Gene/Protein Information For PHB (Source: Uniprot.org, NCBI)
Gene Name
PHB
Full Name
Prohibitin
Weight
29.804kDa
Superfamily
prohibitin family
Alternative Names
Hepatocyte growth factor-regulated tyrosine kinase substrate; Hrs; Protein pp110; HGS; HRS PHB HEL-215, HEL-S-54e1, PHB prohibitin prohibitin|epididymis luminal protein 215|epididymis secretory sperm binding protein Li 54e
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on PHB, check out the PHB Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for PHB: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-Prohibitin/PHB Antibody Picoband® (A01178-1)
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