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49398
Epithelial-Mesenchymal Transition (EMT) IF Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Epithelial-Mesenchymal Transition (EMT) IF Antibody Sampler Kit #49398

Citations (5)
Simple Western™ analysis of lysates (1mg/mL) from MCF-7 cells using ZO-1 (D6L1E) Rabbit mAb #13663. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 66-440 kDa separation module.
Simple Western™ analysis of lysates (0.1 mg/mL) from MCF-7 cells using E-Cadherin (24E10) Rabbit mAb #3195. The virtual lane view (left) shows a single target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Confocal immunofluorescent analysis of fixed frozen mouse kidney labeled with Vimentin (D21H3) XP® Rabbit mAb (left, green) and co-labeled with F4/80 (BM8.1) Rat mAb (right, red), and ProLong® Gold Antifade Reagent with DAPI #8961 (right, blue).
Confocal immunofluorescent analysis of fixed frozen mouse colon labeled with Vimentin (D21H3) XP® Rabbit mAb (left, green) and co-labeled with F4/80 (BM8.1) Rat mAb (right, red), and ProLong® Gold Antifade Reagent with DAPI #8961 (right, blue).
Confocal immunofluorescent analysis of fixed frozen mouse cerebellum labeled with Vimentin (D21H3) XP® Rabbit mAb (left, green) and co-labeled with F4/80 (BM8.1) Rat mAb (right, red), and ProLong® Gold Antifade Reagent with DAPI #8961 (right, blue).
Simple Western™ analysis of lysates (1 mg/mL) from HeLa cells using Vimentin (D21H3) XP ® Rabbit mAb #5741. The virtual lane view (left) shows a single target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Confocal immunofluorescent analysis of fixed frozen mouse cerebellum labeled with Vimentin (D21H3) XP® Rabbit mAb (left, green) and co-labeled with F4/80 (BM8.1) Rat mAb (right, red), and ProLong® Gold Antifade Reagent with DAPI #8961 (right, blue).
Western blot analysis of extracts from CT26.WT, MC38, and LL/2 (LLC1) cells using TWIST1 (E5G9Y) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Confocal immunofluorescent analysis of fixed frozen E12.5 mouse embryo labeled with TWIST1 (E5G9Y) Rabbit mAb (green), DyLight 554 Phalloidin #13054 (red), and ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Confocal immunofluorescent analysis of fixed frozen E12.5 mouse embryo labeled with TWIST1 (E5G9Y) Rabbit mAb (green), DyLight 554 Phalloidin #13054 (red), and ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Confocal immunofluorescent analysis of SH-SY5Y cells (left, positive) and HeLa cells (right, negative) using TWIST1 (E5G9Y) Rabbit mAb (green), DyLight 650 Phalloidin #12956 (red), and DAPI #4083 (blue).
Western blot analysis of extracts from A172 and MCF7 cells using N-Cadherin (D4R1H) XP® Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from various cell lines using ZO-1 (D6L1E) Rabbit mAb (upper) and α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower).
Western blot analysis of extracts from IGROV-1, A549, and 293 cells using Claudin-1 (D3H7C) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from various cell lines, using E-Cadherin (24E10) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human prostate adenocarcinoma using E-Cadherin (24E10) Rabbit mAb performed on the Leica BOND Rx.
Western blot analysis of extracts from various cell lines using Vimentin (D21H3) XP® Rabbit mAb.
Western blot analysis of extracts from various cell lines using ZEB1 (E2G6Y) XP® Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Western blot analysis of extracts from SH-SY5Y, TOV-112D, and HeLa cells using TWIST1 (E5G9Y) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). Low or negative expression of TWIST1 protein in HeLa cells is consistent with the predicted expression pattern.
Western blot analysis of extracts from A204, SKMEL5, and NIH/3T3 cells using Slug (C19G7) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human non-Hodgkin Lymphoma using N-Cadherin (D4R1H) XP® Rabbit mAb performed on the Leica® BOND Rx.
Immunoprecipitation of ZO-1 from MCF7 cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or ZO-1 (D6L1E) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot was performed using ZO-1 (D6L1E) Rabbit mAb.
Confocal immunofluorescent analysis of IGROV-1 (positive, left), A549 (positive, center), and 293 (negative, right) cells using Claudin-1 (D3H7C) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Immunohistochemical analysis of paraffin-embedded human papillary thyroid carcinoma using E-Cadherin (24E10) Rabbit mAb performed on the Leica BOND Rx.
Western blot analysis of extracts from control HeLa cells (lane 1) or Vimentin knockout HeLa cells (lane 2) using Vimentin (D21H3) XP® Rabbit mAb #5741 (upper) or β-Actin (13E5) Rabbit mAb #4970 (lower). The absence of signal in the Vimentin knockout HeLa cells confirms specificity of the antibody for Vimentin.
Immunoprecipitation of ZEB1 protein from Jurkat cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is ZEB1 (E2G6Y) XP® Rabbit mAb. Western blot analysis was performed using ZEB1 (E2G6Y) XP® Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as the secondary antibody.
Immunohistochemical analysis of paraffin-embedded human granulosa cell tumor of the ovary using N-Cadherin (D4R1H) XP® Rabbit mAb performed on the Leica® BOND Rx.
Confocal immunofluorescent analysis of MCF7 (high-expressing, left) and MDA-MB-435 (low-expressing, right) cells, using ZO-1 (D6L1E) Rabbit mAb (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using E-Cadherin (24E10) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human squamous cell lung carcinoma using Vimentin (D21H3) XP® Rabbit mAb performed on the Leica® BOND Rx.
Immunohistochemical analysis of paraffin-embedded human ductal breast carcinoma using ZEB1 (E2G6Y) XP® Rabbit mAb performed on the Leica® BOND Rx.
Immunohistochemical analysis of paraffin-embedded human gastric carcinoma using N-Cadherin (D4R1H) XP® Rabbit mAb performed on the Leica® BOND Rx.
Immunohistochemical analysis of paraffin-embedded human metastatic adenocarcinoma in lymph node, using E-Cadherin (24E10) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using Vimentin (D21H3) XP® Rabbit mAb performed on the Leica® BOND Rx.
Immunohistochemical analysis of paraffin-embedded human mucoepidermoid carcinoma of the larynx using ZEB1 (E2G6Y) XP® Rabbit mAb performed on the Leica® BOND Rx.
Confocal immunofluorescent analysis of A204 cells (left) and PANC-1 cells (right) using Slug (C19G7) Rabbit mAb (green). Actin filaments have been labeled with DY554 phalloidin (red).
Immunohistochemical analysis of paraffin-embedded human colon using N-Cadherin (D4R1H) XP® Rabbit mAb. Note staining of myenteric plexus.
Immunohistochemical analysis of paraffin-embedded mouse prostate using E-Cadherin (24E10) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Vimentin (D21H3) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human ovarian serous carcinoma using ZEB1 (E2G6Y) XP® Rabbit mAb.
Flow cytometric analysis of PANC-1 cells (blue) and A204 cells (green) using Slug (C19G7) Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma using N-Cadherin (D4R1H) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded mouse pancreas using E-Cadherin (24E10) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded mouse colon using Vimentin (D21H3) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using ZEB1 (E2G6Y) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded A172 (positive, left) and MCF7 (negative, right) cell pellets using N-Cadherin (D4R1H) XP® Rabbit mAb.
Confocal immunofluorescent images of MCF7 cells using E-Cadherin (24E10) Rabbit mAb (green, left) compared to an isotype control (right). Blue pseudocolor = DRAQ5® (fluorescent DNA dye).
Immunohistochemical analysis of paraffin-embedded mouse small intestine using E-Cadherin (24E10) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded rat colon using Vimentin (D21H3) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human prostate adenocarcinoma using ZEB1 (E2G6Y) XP® Rabbit mAb.
Confocal immunofluorescent analysis of A172 (positive, left) and MCF7 (negative, right) cells using N-Cadherin (D4R1H) XP® Rabbit mAb (green). Blue pseudocolor= DRAQ5® #4084 (fluorescent DNA dye).
Immunohistochemical analysis of paraffin-embedded mouse lung using E-Cadherin (24E10) Rabbit mAb.
Confocal immunofluorescent analysis of SNB19 cells using Vimentin (D21H3) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Immunohistochemical analysis of paraffin-embedded rhesus kidney using Vimentin (D21H3) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using ZEB1 (E2G6Y) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded mouse stomach using E-Cadherin (24E10) Rabbit mAb.
Flow cytometric analysis of MCF7 cells (blue, negative) and HeLa cells (green, positive) using Vimentin (D21H3) XP® Rabbit mAb(solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Immunohistochemical analysis of paraffin-embedded Syrian hamster small intestine using Vimentin (D21H3) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human gastrointestinal stromal tumor using ZEB1 (E2G6Y) XP® Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using E-Cadherin (24E10) Rabbit mAb in the presence of control peptide (left) or E-Cadherin Blocking Peptide #1050 (right).
Flow cytometric analysis of Jurkat cells (blue, negative) and MCF7 cells (green, positive) using E-Cadherin (24E10) Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')₂ Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Immunohistochemical analysis of paraffin-embedded human tonsil using Vimentin (D21H3) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
Immunohistochemical analysis of paraffin-embedded mouse lung using ZEB1 (E2G6Y) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded 4T1 syngeneic mammary tumor using ZEB1 (E2G6Y) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded HT-1080 cell pellet (left, positive) or HCC1806 cell pellet (right, negative) using ZEB1 (E2G6Y) XP® Rabbit mAb.
Confocal immunofluorescent analysis of HT-1080 cells (left, positive) and HCC1806 cells (right, negative) using ZEB1 (E2G6Y) XP® Rabbit mAb (green), Cytochrome c (6H2.B4) Mouse mAb #12963 (yellow), and Alexa Fluor® 647 Phalloidin #8940 (red). Slides were mounted with ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Flow cytometric analysis of HCC1806 cells (blue, negative) and HT-1080 cells (green, positive) using ZEB1 (E2G6Y) XP® Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
To Purchase # 49398
Cat. # Size Qty. Price
49398T
1 Kit  (8 x 20 microliters)

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Claudin-1 (D3H7C) Rabbit mAb 13995 20 µl
  • WB
  • IF
H 20 Rabbit IgG
E-Cadherin (24E10) Rabbit mAb 3195 20 µl
  • WB
  • IHC
  • IF
  • F
H M 135 Rabbit IgG
N-Cadherin (D4R1H) XP® Rabbit mAb 13116 20 µl
  • WB
  • IP
  • IHC
  • IF
H M 140 Rabbit IgG
Slug (C19G7) Rabbit mAb 9585 20 µl
  • WB
  • IP
  • IF
  • F
H M 30 Rabbit IgG
TWIST1 (E5G9Y) Rabbit mAb 90445 20 µl
  • WB
  • IF
H M 26 Rabbit IgG
Vimentin (D21H3) XP® Rabbit mAb 5741 20 µl
  • WB
  • IHC
  • IF
  • F
H M R Mk 57 Rabbit IgG
ZEB1 (E2G6Y) XP® Rabbit mAb 70512 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
H M R 200 Rabbit IgG
ZO-1 (D6L1E) Rabbit mAb 13663 20 µl
  • WB
  • IP
  • IF
H Mk 220 Rabbit IgG

Product Description

The Epithelial-Mesenchymal Transition (EMT) IF Antibody Sampler Kit provides an economical means to evaluate the expression of established markers of EMT by immunofluorescence (IF).

Specificity / Sensitivity

Claudin-1 (D3H7C) Rabbit mAb detects endogenous levels of total claudin-1 protein. In both claudin-1-positive and claudin-1-negative cell lines, the antibody cross-reacts with a band of unknown origin, which migrates at approximately 120 kDa by SDS-PAGE electrophoresis. This non-specific signal is not detected by fluorescent immunostaining. E-Cadherin (24E10) Rabbit mAb detects endogenous levels of total E-cadherin protein. The antibody does not cross-react with related family members, such as N-cadherin. N-Cadherin (D4R1H) XP® Rabbit mAb detects endogenous levels of total N-cadherin protein. Some non-specific immunostaining has been observed in mouse kidney tissue. Slug (C19G7) Rabbit mAb detects endogenous levels of total Slug protein. TWIST1 (E7E2G) Rabbit mAb (IF Formulated) detects endogenous levels of total TWIST1 protein. Vimentin (D21H3) XP® Rabbit mAb detects endogenous levels of total vimentin protein. ZEB1 (E2G6Y) XP® Rabbit mAb detects endogenous levels of total ZEB1 protein. ZO-1 (D6L1E) Rabbit mAb detects endogenous levels of total ZO-1 protein.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro194 of human claudin-1 protein, Pro780 of human E-cadherin protein, Arg526 of human N-cadherin protein, Pro24 of human TWIST1 protein, Arg45 of human vimentin protein, or Ala1558 of human ZO-1 protein. Monoclonal antibodies are produced by immunizing animals with a recombinant protein corresponding to human Slug protein or the central region of human ZEB1 protein.

Background

Epithelial-mesenchymal transition (EMT) is an essential process during development whereby epithelial cells acquire mesenchymal, fibroblast-like properties and display reduced intracellular adhesion and increased motility. This is a critical feature of normal embryonic development, which is also utilized by malignant epithelial tumors to spread beyond their origin (1-3). This tightly regulated process is associated with a number of cellular and molecular events. EMT depends on a reduction in expression of cell adhesion molecules. Cadherins mediate calcium-dependent cell-cell adhesion and play critical roles in normal tissue development (4). E-cadherin is considered an active suppressor of invasion and growth of many epithelial cancers (4-6). Recent studies indicate that cancer cells have upregulated N-cadherin in addition to loss of E-cadherin. This change in cadherin expression is called the "cadherin switch" and downregulation of E-cadherin is one of the hallmarks of EMT (1). Tight junctions, or zonula occludens, form a continuous barrier to fluids across the epithelium and endothelium. They function in regulation of paracellular permeability and in the maintenance of cell polarity, blocking the movement of transmembrane proteins between the apical and the basolateral cell surfaces. Tight junctions are composed of claudin and occludin proteins, which join the junctions to the cytoskeleton (7,8). Zona occludens (ZO) proteins (e.g., ZO-1) are peripheral membrane adaptor proteins that link junctional transmembrane proteins such as occludin and claudin to the actin cytoskeleton (9). ZO proteins are required for tight junction formation and function (10,11); mutations in ZO-1 and claudin induce EMT (12). Vimentin is an intermediate filament of mesenchymal origin and is present at early developmental stages. Vimentin's dynamic structural changes and spatial re-organization in response to extracellular stimuli helps to coordinate various signaling pathways (13). Slug (SNAI2) is a widely expressed transcriptional repressor and member of the Snail family of zinc finger transcription factors (14,15). Similar to the related Snail protein, Slug binds to the E-cadherin promoter region to repress transcription during development (16). The binding of Slug to integrin promoter sequences represses integrin expression and results in reduced cell adhesion (17). ZEB family proteins (e.g., ZEB1) are zinc finger and homeobox domain containing transcription factors, whose targets of regulation include E-cadherin (1). TWIST1 is a basic helix-loop-helix (b-HLH) transcription factor that functions as a master regulator of embryonic morphogenesis, and plays essential roles in mesenchymal differentiation (18,19). TWIST is upregulated in various human tumors and has been suggested to be a driver of EMT and metastasis (20-21).

  1. Aigner, K. et al. (2007) Oncogene 26, 6979-88.
  2. Peinado, H. et al. (2007) Nat Rev Cancer 7, 415-28.
  3. Moreno-Bueno, G. et al. (2008) Oncogene 27, 6958-69.
  4. Wheelock, M.J. and Johnson, K.R. (2003) Annu Rev Cell Dev Biol 19, 207-35.
  5. Christofori, G. (2003) EMBO J 22, 2318-23.
  6. Hazan, R.B. et al. (2004) Ann N Y Acad Sci 1014, 155-63.
  7. Shin, K. et al. (2006) Annu Rev Cell Dev Biol 22, 207-35.
  8. Oliveira, S.S. and Morgado-Díaz, J.A. (2007) Cell Mol Life Sci 64, 17-28.
  9. Matter, K. and Balda, M.S. (2007) J Cell Sci 120, 1505-11.
  10. Hernandez, S. et al. (2007) Exp Cell Res 313, 1533-47.
  11. Umeda, K. et al. (2006) Cell 126, 741-54.
  12. Reichert, M. et al. (2000) J Biol Chem 275, 9492-500.
  13. Helfand, B.T. et al. (2004) J Cell Sci 117, 133-41.
  14. Inukai, T. et al. (1999) Mol Cell 4, 343-52.
  15. Barrallo-Gimeno, A. and Nieto, M.A. (2005) Development 132, 3151-61.
  16. Bolós, V. et al. (2003) J Cell Sci 116, 499-511.
  17. Turner, F.E. et al. (2006) J Biol Chem 281, 21321-31.
  18. Lee, M.S. et al. (1999) J Cell Biochem 75, 566-77.
  19. Chen, Z.F. and Behringer, R.R. (1995) Genes Dev 9, 686-99.
  20. Yang, J. et al. (2004) Cell 117, 927-39.
  21. Watanabe, O. et al. (2005) Anticancer Res 24, 3851-6.

Pathways

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Limited Uses

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For Research Use Only. Not for Use in Diagnostic Procedures.
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U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
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