Anti-Prohibitin/PHB Antibody

Product Name

Anti-Prohibitin/PHB Antibody

View all Prohibitin Antibodies

SKU/Catalog Number

PA1932

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-Prohibitin/PHB Antibody catalog # PA1932. Tested in IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.

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 (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1932)

Host

Rabbit

Contents

Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg Thimerosal, 0.05mg NaN3.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

A synthetic peptide corresponding to a sequence at the C-terminus of human Prohibitin, identical to the related rat and mouse sequences.

*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

PA1932 is reactive to PHB in Human, Mouse, Rat

Applications

PA1932 is guaranteed for IF, IHC, ICC, WB Boster Guarantee

Observed Molecular Weight

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.

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Reconsitution

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

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.

Immunocytochemistry/Immunofluorescence, 2μg/ml, Human, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, Rat, Mouse, By Heat
Western blot, 0.1-0.5μg/ml, Human, Mouse, Rat

Validation Images & Assay Conditions

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Anti-Prohibitin antibody, PA1932, Western blotting
Lane 1: Rat Lung Tissue Lysate
Lane 2: Rat Skeletal Muscle Tissue Lysate
Lane 3: Rat Brain Tissue Lysate
Lane 4: Rat Kidney Tissue Lysate
Lane 5: HELA Cell Lysate
Lane 6: MCF-7 Cell Lysate
Lane 7: PC-12 Cell Lysate
Lane 8: A549 Cell Lysate
Lane 9: SMMC Cell Lysate

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Anti-Prohibitin antibody, PA1932, IHC(P)
IHC(P): Human Lung Cancer Tissue

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Anti-Prohibitin antibody, PA1932, ICC
ICC: HELA Cell

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Figure 4. IHC analysis of Prohibitin using anti-Prohibitin antibody (PA1932).
Prohibitin was detected in paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Prohibitin Antibody (PA1932) 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.

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Figure 5. Western blot analysis of Prohibitin using anti-Prohibitin antibody (PA1932).
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 50ug of sample under reducing conditions.
Lane 1: mouse NIH3T3 whole cell lysate.
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 antigen affinity purified polyclonal antibody (Catalog # PA1932) 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 1:10000 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 at approximately 30KD. The expected band size for Prohibitin is at 30KD.

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Figure 6. IF analysis of Prohibitin using anti-Prohibitin antibody (PA1932).
Prohibitin was detected in immunocytochemical section of U20S cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL rabbit anti-Prohibitin Antibody (PA1932) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Gene/Protein Information For PHB (Source: Uniprot.org, NCBI)

Alternative Names

PHB; PHB1; Prohibitin PHB HEL-215, HEL-S-54e1, PHB prohibitin prohibitin|epididymis luminal protein 215|epididymis secretory sperm binding protein Li 54e

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].

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