- 35-2900 - Invitrogen Antibodies EPHB4 antibody

Submitted references Class I and III HDACs and loss of active chromatin features contribute to epigenetic silencing of CDX1 and EPHB tumor suppressor genes in colorectal cancer.

Rönsch K, Jäger M, Schöpflin A, Danciu M, Lassmann S, Hecht A
Epigenetics 2011 May;6(5):610-22

Class I and III HDACs and loss of active chromatin features contribute to epigenetic silencing of CDX1 and EPHB tumor suppressor genes in colorectal cancer.

Rönsch K, Jäger M, Schöpflin A, Danciu M, Lassmann S, Hecht A
Epigenetics 2011 May;6(5):610-22

The EphA4 receptor regulates neuronal morphology through SPAR-mediated inactivation of Rap GTPases.

Richter M, Murai KK, Bourgin C, Pak DT, Pasquale EB
The Journal of neuroscience : the official journal of the Society for Neuroscience 2007 Dec 19;27(51):14205-15

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of EphB4 (green) in HT29 cells. The cells were fixed with 4% paraformaldehyde in PBS for 15 minutes at room temperature and blocked with 3% BSA for 30 minutes at room temperature. Cells were stained with a EphB4 mouse monoclonal antibody (Product # 35-2900) at a concentration of 5 µg/mL in blocking buffer for 1 hour at room temperature, and then incubated with a Goat anti-Mouse IgG (H+L) Secondary Antibody, Alexa Fluor Plus 488 conjugate (Product # A32731) at a dilution of 1:500 for at least 30 minutes at a room temperature in the dark (green). Nuclei (blue) were stained with Hoechst 33342 (Product # 62249). Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of EphB4 was performed using 70% confluent log phase HT-29, A549 cells. 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 1 hour at room temperature. The cells were labeled with EphB4 Mouse Monoclonal Antibody (3D7F8) (Product # 35-2900) at 4 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant 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 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 Cytoplasmic localization. Panel e represents A549 cells having no expression of EphB4. 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: Immunohistochemistry analysis of EPHB4 RECEPTOR showing staining in the cytoplasm of paraffin-embedded human kidney tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a EPHB4 RECEPTOR Mouse Monoclonal Antibody (Product # 35-2900) diluted in 3% BSA-PBS at a dilution of 1:20 for 1 hour at 37ºC in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

35-2900

Antibody data