35-2900

antibody from Invitrogen Antibodies

Targeting: EPHB4 HTK, Tyro11

Western blot (Recommended by provider)

ELISA (Recommended by provider)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Data presented on provider website)

Antibody Data

Product number

35-2900

Provider

Invitrogen Antibodies

Product name

EphB4 Monoclonal Antibody (3D7F8)

Antibody type

Monoclonal

Antigen

Recombinant full-length protein

Description

37-1800 has been successfully used in Western Blot, immunohistochemical, immunofluorescence, and ELISA assays. 35-2900 by western blot detects a ~125kDa band representing EphB4 but also detects a strong band at ~90kDa , likely representing another protein in the ephrin family. 37-1800 is recommended for WB.

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

3D7F8

Vial size

100 µg

Concentration

0.5 mg/mL

Storage

-20°C

References

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

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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

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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

Immunocytochemistry

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.