| Size | Price | Stock | Qty |
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| 10mg |
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| 50mg |
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| 100mg |
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| Other Sizes |
| Targets |
Phytohemagglutinin P targets carbohydrate structures, specifically complex N-linked glycans containing galactose, N-acetylgalactosamine, and mannose residues on cell surface glycoproteins and glycolipids. The primary cellular target is the T cell receptor-associated CD3 complex and other signaling molecules, but the mechanism involves cross-linking of multiple glycoproteins leading to global T cell activation. By binding to these targets, PHA-P bypasses antigen specificity and activates a large fraction of T cells via the TCR/CD3 pathway.
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| ln Vitro |
Phytohemagglutinin-P (10 μg/ml; 24-48hous)-stimulated decidual CD8+ T lymphocytes produces high levels of both interferon gamma and interleukin (IL) 8, and low levels of granulocyte-macrophage colony-stimulating factor (CSF2), IL1B, IL2, IL6, IL10, IL12, and tumor necrosis factor; these did not vary with gestational age[2].
In vitro, Phytohemagglutinin P exhibits potent mitogenic activity. At concentrations of 1-10 microg/mL, it stimulates human peripheral blood mononuclear cells (PBMCs) to undergo blast transformation, upregulate activation markers (CD25, CD69, HLA-DR), and proliferate. The peak of DNA synthesis occurs at 48-72 hours as measured by [3H]-thymidine incorporation or BrdU uptake. PHA-P also induces cytokine production, especially IL-2, IFN-gamma, and TNF-alpha. In addition, it causes agglutination of red blood cells and leukocytes due to its hemagglutinating activity. |
| ln Vivo |
Phytohemagglutinin-P (25 and 50 mg/kg; ip; daily for 15 days; male Sprague-Dawley rats) shows a decrease in longitudinal bone growth, in the number of chondrocytes, in the thickness of cartilage plate, in the metaphyseal mass of hard tissue, in the percentage of calcified cartilage core, and in the number of osteoblasts per mm of bone surface. Additionally, PHA-P increases the number of osteoclasts, the number of labeled osteoclastic nuclei, and the average number of nuclei per osteoclast[ 1].
Phytohemagglutinin P is not typically used as a systemic therapeutic agent in vivo due to its potent immunostimulatory and toxic effects. However, it has been studied in animal models as an immune adjuvant. When injected subcutaneously or intraperitoneally into mice, PHA-P induces local inflammation, splenomegaly, and increased serum cytokine levels. It can also cause lethal systemic inflammatory responses at high doses. Topical application of PHA-P has been explored for treating cutaneous viral infections, but this is not standard practice. In vivo, the compound is rapidly cleared and may cause anaphylaxis. |
| Enzyme Assay |
A non-cellular (cell-free) experiment for PHA-P is its hemagglutination assay. A suspension of washed red blood cells (human or rabbit) at 2% concentration is mixed with serially diluted PHA-P solution in round-bottom 96-well plates. After 1-2 hours at room temperature, the pattern of settled cells is observed. A diffuse button indicates no agglutination, while a lattice covering the bottom indicates agglutination. The minimum agglutinating concentration (MAC) is determined as the highest dilution that still causes complete agglutination. This assay measures the lectin's carbohydrate-binding activity.
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| Cell Assay |
Cellular in vitro experiments are performed using isolated human PBMCs or purified T cells. Cells are seeded in 96-well plates at 1-2 x 10⁵ cells/well in RPMI-1640 medium with 10% fetal bovine serum. PHA-P is added at final concentrations of 0.5-20 microg/mL. After 48-72 hours of incubation at 37degC in 5% CO2, cell proliferation is measured by adding 0.5 microCi of [3H]-thymidine per well for the last 4-8 hours, followed by cell harvesting and scintillation counting. Alternatively, a colorimetric MTT or CellTiter-Glo assay can be used. Supernatants can be collected for cytokine ELISA.
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| Animal Protocol |
In vivo animal experiments with PHA-P are limited due to toxicity. In a typical protocol, female BALB/c mice are injected intraperitoneally with PHA-P (50-500 microg/mouse). After 2-6 hours, blood is collected, and serum cytokines (IL-2, IL-6, TNF-alpha) are measured by ELISA. Spleens are harvested for flow cytometry analysis of T cell activation markers and for ex vivo lymphocyte proliferation assays. At higher doses (>1 mg/mouse), animals show signs of septic shock (hypothermia, piloerection, lethargy) and may die within 24 hours. Doses for adjuvant studies are typically much lower (10-50 microg/mouse).
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| ADME/Pharmacokinetics |
Phytohemagglutinin P is a protein and would be digested if given orally, so it is not administered by that route. When injected intravenously or intraperitoneally, it has a short plasma half-life (minutes to hours) due to rapid clearance by the reticuloendothelial system and proteolytic degradation. It is not absorbed through intact skin. Its distribution is largely restricted to the vascular space and lymphoid organs. Metabolism involves proteolysis in the liver and other tissues. Excretion occurs via urine as small peptide fragments. No formal PK studies have been published for PHA-P as it is not a drug candidate.
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| Toxicity/Toxicokinetics |
Phytohemagglutinin P is toxic at high doses. The LD50 in mice after intravenous injection is approximately 5-10 mg/kg. Symptoms include respiratory distress, diarrhea, and hypothermia. Chronic exposure can cause weight loss, lymphoid hyperplasia, and autoimmune-like phenomena. Occupational exposure to dust from kidney beans can cause respiratory sensitization. PHA-P is also a potent inducer of histamine release from basophils and can cause anaphylaxis if injected. Extreme caution is required when handling the pure powder; use gloves, mask, and work in a biosafety cabinet.
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| References | |
| Additional Infomation |
Phytohemagglutinin P is extracted from red kidney beans and purified by affinity chromatography. It is supplied as a lyophilized powder, soluble in water and buffers. One of the main applications is in karyotyping: PHA-P stimulates T lymphocytes in whole blood to divide, allowing metaphase arrest and chromosome preparation for prenatal and postnatal genetic analysis. It is also used in cancer immunology to generate lymphokine-activated killer (LAK) cells and in HIV research to activate latent virus. PHA-P is not an approved drug; it is a research-grade reagent with no clinical indications.
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| Appearance |
Solid powder
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| Synonyms |
Phytohemagglutinin P; PHA-P
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: This product requires protection from light (avoid light exposure) during transportation and storage. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
H2O : 10 mg/mL (with sonication and heat)
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| Solubility (In Vivo) |
Solubility in Formulation 1: 25 mg/mL in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution sonicated
(Please use freshly prepared in vivo formulations for optimal results.) |
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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