Guidelines on the biological function and application of phytohemagglutinin

Product introduction:

 

Phytohemagglutinin (PHA) refers to a bean lectin extracted from red kidney bean (Phaseolus Vulgaris). It is a tetrameric glycoprotein formed by non-covalent bonding of 4 subunits, containing two subunit molecules, L subunit (leukoagglutinin) and E (erythroagglutinin), containing 5 isomers (L4, L3E1, L2E2, L1E3 and E4). The L subunit has leukocyte agglutination and high mitogenic activity; The E subunit has high erythrocyte agglutination and low mitogenic activity.

 

According to different subunit compositions, PHA can be divided into several different forms, mainly including PHA-L, PHA-E, PHA-P and PHA-M. The following are their main uses in scientific research:

 

PHA-L:

 

Main function: PHA-L is a purified L4 tetramer, mainly as a stimulator of T lymphocytes, and is widely used in immunological research. It can be used as a positive control for INF-γICS, ELISPOT experiments; Stimuli of PBMC culture; Anterograde nerve tracers, etc.

Biological activity: It has high mitogenic activity and leukocyte agglutination activity, but extremely low erythrocyte agglutination activity.

Application: It is mainly used to stimulate T cell proliferation and immune response research.

 

PHA-E:

 

Main function: PHA-E is a purified E4 tetramer, and its main use is the study of erythrocyte agglutination or glycosylation modification.

Biological activity: It has high erythrocyte agglutination activity and low mitogenic activity.

Application: It is mainly used for erythrocyte agglutination experiments and glycosylation studies.

 

PHA-P:

 

Main function: PHA-P is the protein form of PHA before the separation and purification of PHA-L and PHA-E, and its composition is comparable to PHA-M.

Biological activity: It has the activity of promoting mitosis and inducing interferon secretion.

Application: It is mainly used to stimulate the proliferation of peripheral mononuclear cells, promote the production of certain cytokines and the expression of membrane surface proteins.

 

PHA-M

 

Main Function: PHA-M is the viscous protein form of phytohemagglutinin, and its composition is comparable to PHA-P. It is mainly used to stimulate the proliferation of peripheral mononuclear cells, promote the production of certain cytokines and the expression of membrane surface proteins.

Biological activity: It has the activity of promoting mitosis and inducing interferon secretion.

Application: It is widely used in immunological research, especially in cell proliferation and cytokine production.

 

Customer application examples:

 

Application 1.

Use PHA-L (abs47014909) to pre-stimulate Jurkat T cells to explore the mechanism of small molecule PROTAC-mediated targeted protein degradation in the treatment of STAT3-dependent epithelial cancer [1]

 

HCT116 cells were seeded at a density of 1 × 104 cells/well into 12-well plates for attachment, followed by treatment with TSM-1 (1 μM) for 24 h. Then, Jurkat T cells (6 × 104) were pre-stimulated with PHA (10 μg/mL) and PMA (10 ng/mL) for 4 h. HCT116 cells were co-cultured with activated Jurkat T cells for 24 h. The ELISA kit was used to detect the level of IL-2 in the supernatant medium, which is a biomarker for monitoring T cell activation. Virtualization of potentially cancerous viable cells by crystal violet staining. After lysing the cells in 10% acetic acid, the cells were quantified using a Cytation 5 microplate reader (BioTek) at 595 nm.

 

Figure 1 TSM-1 induces T cell activation and infiltration in vitro

 

Application 2:

Jurkat T cells were cultured using PHA-M (abs47014911) to explore the mechanism of fluoride interfering with the immune system [2].

 

Jurkat T cells (HTX4056, Otwo Biotech) were grown in RPMI 1640 medium supplemented with 10% fetal bovine serum (FBS, abs972, Absin), penicillin-streptomycin solution and 5 ug/mL phytohemagglutinin-M (abs47014911, Absin), 5 ug/mL Ultra LEAFTM purified anti-human CD28. Cell lines were cultured in a humidified incubator at 37 °C, 5% CO2 and 95% air.

 

Fig. 2 Fluoride can inhibit T cell activation and hinder the internalization of T cell receptors

 

Different forms of PHA mainly differ in their subunit composition and corresponding biological functions. PHA-L and PHA-E have high mitogenic activity and high erythrocyte agglutination activity, respectively, while PHA-M and PHA-P are mainly used to stimulate cell proliferation and cytokine production. Each form has its specific applications in immunological research and cell biology research.

 

References

[1] Jin J, Wu Y, Zhao Z, et al. Small-molecule PROTAC mediates targeted protein degradation to treat STAT3-dependent epithelial cancer [J]. JCI Insight. 2022; 7 (22): e160606.

[2] Zhu S, Liu B, Fu G, et al. PKC-θ is an important driver of fluoride-induced immune imbalance of regulatory T cells/effect T cells [J]. Sci Total Environ. 2024;934:173081.

 

PHA product recommendations:

Item number	Product name	Specifications
abs47014910	Phytohemagglutinin-L solution (500 ×)	100uL
abs47014909	Phytohemagglutinin-L (Phaseolus vulgaris)	5mg
abs47014911	Phytohemagglutinin-M (Phaseolus vulgaris)	5mg
abs47014912	Phytohemagglutinin-E (Phaseolus vulgaris)	5mg
abs47014913	Phytohemagglutinin-P (Phaseolus vulgaris)	5mg

 

Related product recommendations:

Item number	Product name	Specifications
abs9107	Phorbol ester (PMA)	5mg
abs551102	Human IL-2 ELISA Kit (Plus)	96 T
abs972	Fetal bovine serum (superior grade)	500mL
abs9484	RPMI 1640 medium (with L-glutamine, without HEPES)	500 mL × 10
abs9244	Penicillin-streptomycin solution (100 ×, double antibody)	100mL

 

Absin provides antibodies, proteins, ELISA kits, cell culture, detection kits, and other research reagents. If you have any product needs, please contact us.

Absin Bioscience Inc. Email: worldwide@absin.cn

Follow us on Facebook: Absin Bio