- MA1-16519 - Invitrogen Antibodies EPAS1 antibody

Submitted references Differential but Complementary HIF1α and HIF2α Transcriptional Regulation.

Downes NL, Laham-Karam N, Kaikkonen MU, Ylä-Herttuala S
Molecular therapy : the journal of the American Society of Gene Therapy 2018 Jul 5;26(7):1735-1745

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot of HIF-2 alpha detected in hypoxic human lysate using Product # MA1-16519. Lane 1: normoxic A549 lysate control, lane 2: hypoxic A549 lysate.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of HIF-2 alpha in HOS cells following exposure to hypoxia. Samples were incubated in HIF-2 alpha monoclonal antibody (Product # MA1-16519). Image using the HRP form of this antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of HIF-2 alpha in 0.5 mg/mL Hypoxic HeLa lysate. Samples were incubated in HIF-2 alpha monoclonal (Product # MA1-16519). This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of HIF-2 alpha in HepG2. Samples were incubated in HIF-2 alpha monoclonal antibody (Product # MA1-16519 using a dilution of 1 - 2 µg/mL. HepG2 without Cobalt (II) Chloride (1), HepG2 with Cobalt (II) Chloride (2), HepG2 normoxic (3), HepG2 hypoxic (4), HepG2 without Cobalt (II) Chloride (5), HepG2 with Cobalt (II) Chloride (6), HepG2 normoxic (7), and HepG2 hypoxic (8).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of HIF-2 alpha in HOS cells following exposure to hypoxia. Samples were incubated in HIF-2 alpha monoclonal antibody (Product # MA1-16519). Image using the HRP form of this antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of HIF-2 alpha in 791T cells following exposure to hypoxia. Samples were incubated in HIF-2 alpha monoclonal antibody (Product # MA1-16519). Image using the HRP form of this antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of HIF-2 alpha in 0.5 mg/mL Hypoxic HeLa lysate. Samples were incubated in HIF-2 alpha monoclonal (Product # MA1-16519). This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of HIF-2 alpha was performed using 70% confluent log phase MCF7 treated with Cobalt Chloride (150 µM for 48hrs). The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with HIF-2 alpha Monoclonal Antibody (ep190b) (Product # MA1-16519) at 2 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32766), (1:2,000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear translocation of HIF-2 alpha upon cobalt chloride treatment in MCF7 cells. Panel e represents untreated MCF7 cells with no signal. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of HIF-2 alpha was performed using 70% confluent log phase HeLa cells treated with 150µM of Cobalt chloride for 48 hours. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with HIF-2 alpha Mouse Monoclonal Antibody (ep190b) (Product # MA1-16519) at 5 microgram/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e shows untreated cells with cytoplasmic signal. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemical analysis of HIF-2 alpha in human cardiac myocytes. Samples were incubated in HIF-2 alpha monoclonal antibody (Product # MA1-16519).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry of HIF-2 alpha in HepG2 cells. Samples were incubated in HIF-2 alpha monoclonal antibody (Product # MA1-16519) using a dilution of 1:400 followed by a Alexa Fluor 488 secondary (shown in purple). M1 is defined by unstained cells.

MA1-16519

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