mAbs Against HUMAN Protein
Leucocyte specific protein 1
| STATUS: |  |
|---|---|
| CONTACT INFORMATION: | LRF Haemato-oncology Group, University of Oxford |
| STATUS: | Validated |
| TYPE: | mouse anti human |
| CLONE NAME: | TPD153 |
| PROTEIN: | LSP-1 |
| ANTIGEN USED: | Tonsil cells |
| FUSION PARTNER: | NS1 |
| ISOTYPE: | IgG1 |
| SPECIES REACTIVITY: | human |
| PREPARATION AND STORAGE: | Aliquot and store at 4ºC. Do not freeze |
| APP RECOMMENDED: | IHQ-frozen, IHQ-paraffin, IF, WB, IP, Flow cytometry |
Description
The antibody detects an intracellular antigen in peripheral blood B cells, monocytes and granulocytes and the 85-90% circulating T cells. Medullary but not cortical thymocytes express LSP1. Plasma cells, dendritic cells and Langerhans cells were also LSP1+ve. The protein was detected in a wide range of leukaemias and non-Hodgkin s lymphomas and the tumour cells in classical Hodgkin \'s lymphoma. Reciprocal expression was noted between LSP1 and ALK expression in ALCL. A combination of LSP1 and anti-CD45 may be useful for identifying haematologucal malignancies which have decreased CD45 expression. LSP1 expression is also of use for the diagnosis of T-cell rich B-cell lymphoma from lymphocyte-rich Hodgkin s lymphoma.
References
Marafioti T, Mancini C, Ascani S, Sabattini E, Zinzani PL, Pozzobon M, Pulford K, Falini B, Jaffe ES, Müller-Hermelink HK, Mason DY, Pileri SA. Leukocyte-specific phosphoprotein-1 and PU.1: two useful markers for distinguishing T-cell-rich B-cell lymphoma from lymphocyte-predominant Hodgkin \'s disease.
Haematologica. 2004 Aug;89(8):957-64.
Marafioti T, Jabri L, Pulford K, Brousset P, Mason DY, Delsol G
Leucocyte-specific protein (LSP1) in malignant lymphoma and Hodgkin s disease
Br J Haematol. 2003 Feb;120(4):671-8
Pulford K, Jones M, Banham AH, Haralambieva E, Mason DY
Lymphocyte-specific protein 1: a specific marker of human leucocytes.
Immunology. 1999 Feb;96(2):262-71
Applications
| IHC Techniques | Clone | Dilution | Antibody concentration | Antigen retrieval method | Visualization kit | +/- control | Protein localization | Positivity in other species | Protocol |
|---|---|---|---|---|---|---|---|---|---|
| Frozen tissue and cytospins | |||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | TPD153 | Neat | Supernantant | Indirect immunoperoxidase or APAAP | anti-BCL2 / Mouse IgG1 | Intracellular | Not known | ||
| Paraffin tissue | |||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | TPD153 | 1/5 | Supernantant | Envision | As above / Universal negative | Intracellular | |||
| Immunofluorescence | |||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | TPD153 | 1/10 | Supernantant | Directly conjugated antibody | As above / As above | Intracellular | |||
Enlarge image
- Antibody TPD153 in tansfectants
- Antibody TPD153 stained LSP1 transfectants. Inset is vector only control
Enlarge image
- Immunofluorescent labelling of thymus to show LSP1 expression.
- LSP1 (green) is expressed in the CD3-positive (red) cells preset in the medulla while only a minority of cortical CD3-positive cells express LSP1 (arrow).
| WB Techniques | Clone | Dilution | Antibody concentration | +/- control | Expected MW | Observed Mw | Positivity in other species | Protocol |
|---|---|---|---|---|---|---|---|---|
| Western Blotting | ||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | TPD153 | Neat | Supernatant | anti-BCL2 / Mouse IgG1 | 60 kD | 60 kD | ||
| Immunoprecipitation | ||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | TPD153 | As above | As above | As above / As above | 60 kD | 60 kD | ||
Enlarge image
- Western blotting studies on LSP1
- a) Antibody TPD153 recognises LSP1 transfectants (pAB280) but not the empty vector (pBK-CMV).
b) Presence of the 60 kD LSP1 protein in tonsil and a range of haematologically-derived cell lines.
| FLOW CYTOMETRY | Clone | Dilution | +/- control | Type of fluorocrom | Protocol |
|---|---|---|---|---|---|
Recommended Result obtained is satisfactory. The reagent can be use in this application | TPD513 | 1/10 | Anti-BCL2 / Mouse IgG1 | FITC |
Enlarge image
- Flow cytometry of the LSP1 protein on tonsil and blood cells.
- a) LSP1 protein is undetectable in unpermeabilised tonsil cells but expressed in b) more than 95% of permeabilised tonsil cells. c), d), e) and f) show the presence of LSP1 protein in both single and double labelling of permeabilsed peripheral blood cells.