MA3-005

antibody from Invitrogen Antibodies

Targeting: EDN1 ET1

Provider product page for MA3-005

Western blot

Immunocytochemistry

Immunohistochemistry

Flow cytometry

Radioimmunoassay

Other assay

Antibody data

Antibody Data
Antigen structure
References [41]
Comments [0]
Validations
Immunocytochemistry [3]
Flow cytometry [3]
Other assay [4]

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Product number: MA3-005 - Provider product page
Provider: Invitrogen Antibodies
Product name: Endothelin 1 Monoclonal Antibody (TR.ET.48.5)
Antibody type: Monoclonal
Antigen: Other
Description: MA3-005 detects ET-1 from human, sheep, mouse and rat tissues. This antibody shows little cross-reactivity to related peptides and no known cross-reactivity to non-related peptides. MA3-005 has been successfully used in Western blot, flow cytometry, immunofluorescence, immunohistochemistry, immunocytochemistry, and radioimmune assay procedures. Immunohistochemical staining of ET-1 in human corpus cavernosum tissue with MA3-005 results in staining of endothelial cells. Radioimmune assays using MA3-005 can be used to concentrate ET-1 in solution (e.g. serµm/plasma, milk, urine). This antibody is not recommended for human placenta tissue or HepG2 cells in Western blot applications. For immunohistochemistry with MA3-005 formalin-fixed, paraffin-embedded tissue sections has been used. The MA3-005 antigen is purified human endothelin-1. Studies suggest that this antibody binds to an epitope in the region of ET-1 represented by amino acids 8-16.
Reactivity: Human, Mouse, Rat
Host: Mouse
Isotype: IgG
Antibody clone number: TR.ET.48.5
Vial size: 100 µL
Concentration: 2.5 mg/mL
Storage: -20° C, Avoid Freeze/Thaw Cycles

EDN1 protein structure - MA3-005 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Endothelin 1 (green) showing positive staining in the secretion of Hela cells (right) compared with a negative control in the absence of primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes, blocked with 3% BSA-PBS for 30 minutes at room temperature and probed with an Endothelin 1 monoclonal antibody (Product # MA3-005) in 3% BSA-PBS at a dilution of 1:200 and incubated overnight at 4 ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight 488-conjugated goat-anti-mouse IgG (H+L) secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with DAPI for 5-10 minutes in the dark. Images were taken at a magnification of 60x.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Endothelin 1 (green) showing positive staining in the secretion of HUVEC cells (right) compared with a negative control in the absence of primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes, blocked with 3% BSA-PBS for 30 minutes at room temperature and probed with an Endothelin 1 monoclonal antibody (Product # MA3-005) in 3% BSA-PBS at a dilution of 1:200 and incubated overnight at 4 ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight 488-conjugated goat-anti-mouse IgG (H+L) secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with DAPI for 5-10 minutes in the dark. Images were taken at a magnification of 60x.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Endothelin 1 (green) showing positive staining in the secretion of PC12 cells (right) compared with a negative control in the absence of primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes, blocked with 3% BSA-PBS for 30 minutes at room temperature and probed with an Endothelin 1 monoclonal antibody (Product # MA3-005) in 3% BSA-PBS at a dilution of 1:200 and incubated overnight at 4 ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight 488-conjugated goat-anti-mouse IgG (H+L) secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with DAPI for 5-10 minutes in the dark. Images were taken at a magnification of 60x.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of Endothelin 1 in 293T cells compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/mL, fixed with 2% paraformaldehyde, washed with PBS, and incubated with Endothelin 1 monoclonal antibody (Product # MA3-005) at a dilution of 0.5 µg/test for 60 min at room temperature. Cells were then blocked in a solution of 2% BSA-PBS for 30 min at room temperature, incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody, and re-suspended in PBS for FACS analysis.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of Endothelin 1 in 3T3 cells compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/mL, fixed with 2% paraformaldehyde, washed with PBS, and incubated with Endothelin 1 monoclonal antibody (Product # MA3-005) at a dilution of 0.5 µg/test for 60 min at room temperature. Cells were then blocked in a solution of 2% BSA-PBS for 30 min at room temperature, incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody, and re-suspended in PBS for FACS analysis.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of Endothelin 1 in HepG2 cells compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/mL, fixed with 2% paraformaldehyde, washed with PBS, and incubated with Endothelin 1 monoclonal antibody (Product # MA3-005) at a dilution of 0.5 µg/test for 60 min at room temperature. Cells were then blocked in a solution of 2% BSA-PBS for 30 min at room temperature, incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody, and re-suspended in PBS for FACS analysis.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: 2 Staining intensities of endothelial cells. Boxplot graphic of iNOS, eNOS, TNF-alpha and ET-1 and MCP-1. Group 1, OVX; Group 2, OVX+RL; Group 3, OVX+AV; Group 4, OVX+RL+AV. Data are presented as median. Kruskal-Wallis test was performed to determine the differences between groups.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 5 Knocking down p38 MAPK protects against endothelial destruction induced by OGD/R. (A) Western blot of phospho-p38 MAPK, p38 MAPK, AQP1, ICAM-1, ZO-1, and VE-cadherin in rPMVECs. (B-F) Quantification of proteins from western blots in panel A. (G) Western blot of eNOS and ET-1 in rPMVECs. (H,I) Quantification of proteins from western blots in panel G. Each experiment was performed independently at least three times. ns, not significant, * p < 0.05.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 5. A, Postoperative evolution of serum ET-1, a marker of endothelial-cell stress and hypoxia. Here, data are expressed as the mean and the standard error of the mean as whiskers, whereas they are expressed as the median and the 25&percnt to 75&percnt interquartile range in the text.&ast P < 0.05 for both SFS and S12 versus S6. B, ET-1 immunostaining of postreperfusion tissue samples. ET-1 expression was significantly upregulated in both the SFS and S12 groups, though the hepatocyte cords remained intact in the latter. S6 grafts, on the other hand, were notable for very minimal ET-1 staining of the endothelial lining.