Human/Mouse PTEN Antibody

Recombinant Monoclonal Antibody
Catalog # Availability Size / Price Qty
MAB847R-100
MAB847R-SP
Detection of Mouse PTEN by Western Blot.
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Human/Mouse PTEN Antibody Summary

Species Reactivity
Human, Mouse
Specificity
Detects human PTEN in direct ELISAs.
Source
Recombinant Monoclonal Mouse IgG1 Clone # 217702R
Purification
Protein A or G purified from cell culture supernatant
Immunogen
E. coli-derived recombinant human PTEN
Thr2-Val403
Accession # P60484
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
2 µg/mL
3T3‑L1 mouse embryonic fibroblast adipose-like cell line, C2C12 mouse myoblast cell line and mouse brain
Intracellular Staining by Flow Cytometry
0.25 µg/106 cells
Human U937 histiocytic lymphoma cell line
Knockout Validated
PTEN is specifically detected in HeLa human cervical epithelial carcinoma parental cell line but is not detectable in PTEN knockout HeLa human cervical epithelial carcinoma cell line.
 

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot View Larger

Detection of Mouse PTEN by Western Blot. Western blot shows lysates of 3T3‑L1 mouse embryonic fibroblast adipose-like cell line and C2C12 mouse myoblast cell line. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human/Mouse PTEN Monoclonal Antibody (Catalog # MAB847R) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for PTEN at approximately 54 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Western Blot View Larger

Detection of Mouse PTEN by Western Blot. Western blot shows lysates of mouse brain. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human/Mouse PTEN Monoclonal Antibody (Catalog # MAB847R) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for PTEN at approximately 54 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Intracellular Staining by Flow Cytometry View Larger

Detection of PTEN in Human U937 cell line by Flow Cytometry. Human U937 histiocytic lymphoma cell line was stained with Mouse Anti-Human/Mouse PTEN Monoclonal Antibody (Catalog # MAB847R, filled histogram) or isotype control antibody (MAB002, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG F(ab')2 Secondary Antibody (F0102B). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (FC005). Staining was performed using our protocol for Staining Intracellular Molecules.

Knockout Validated View Larger

Western Blot Shows Human PTEN Specificity by Using Knockout Cell Line. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line and human PTEN knockout cell line. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human/Mouse PTEN Monoclonal Antibody (Catalog # MAB847R) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for PTEN at approximately 54 kDa (as indicated) in the parental HeLa human cervical epithelial carcinoma cell line, but is not detectable in knockout HeLa human cervical epithelial carcinoma cell line. GAPDH (Catalog # MAB5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Reconstitution Calculator

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS.
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Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: PTEN

The tumor suppressor gene PTEN (phosphatase and tensin homolog deleted on chromosome 10), also known as MMAC1 (mutated in multiple advanced cancers 1), encodes a phosphatase that contains the catalytic signature motif (HCxxGxxRS/T) found in all members of the protein tyrosine phosphatase family. In vitro, the recombinant PTEN has both lipid phosphatase and protein phosphatase activities (1, 2). Interestingly, accumulating evidence has shown that the tumor suppressor activity of PTEN relies on its ability to dephosphorylate phosphatidylinositol (3,4,5)-triphosphate specifically at position 3 of the inositol ring (3). This activity reduces the levels of phosphatidylinositol (3,4,5)-triphosphate which is specifically produced from phosphatidylinositol (4,5)-diphosphate by PI 3-kinase upon activation by a variety of stimuli. Therefore, PTEN antagonizes PI 3-kinase-induced downstream signaling events and cellular processes including cell growth, apoptosis and cell motility. In vivo, the importance of PTEN catalytic activity in its tumor suppressor functions is underscored by the fact that the majority of PTEN missense mutations detected in tumor specimens target the phosphatase domain and cause a loss in PTEN phosphatase activity (4).

References
  1. Maehama, T. and J. Dixon (1998) J. Biol. Chem. 273:13375.
  2. Das, S. et al. (2003) Proc. Natl. Acad. Sci. USA 100:7491.
  3. Myers, M. et al. (1998) Proc. Natl. Acad. Sci. USA 95:13513.
  4. Waite, K. and C. Eng (2002) Am. J. Hum. Genet. 70:829.
Long Name
Phosphatase and Tensin Homolog Deleted on Chromosome 10
Entrez Gene IDs
5728 (Human); 19211 (Mouse); 50557 (Rat)
Alternate Names
BZS; EC 3.1.3.16,10q23del; EC 3.1.3.48; EC 3.1.3.67; GLM2; MMAC1 phosphatase and tensin homolog deleted on chromosome 10; MMAC1; MMAC1MGC11227; Mutated in multiple advanced cancers 1; phosphatase and tensin homologDEC; phosphatidylinositol-34,5-trisphosphate 3-phosphatase and dual-specificityprotein phosphatase PTEN; PTEN; PTEN1; TEP1MHAM

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