Anti-Eph receptor B1 + Eph receptor B2 (phospho Y594 + Y596) antibody

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Eph receptor B1 + Eph receptor B2 (phospho Y594 + Y596) antibody (AB61791)

Immunofluorescence analysis of HuvEc cells, using ab61791 at 1/500-1/1000 dilution with (+) or without (-) immunising phosphopeptide.

• ICC/IF

AbReview41906****

Immunocytochemistry/ Immunofluorescence - Anti-Eph receptor B1 + Eph receptor B2 (phospho Y594 + Y596) antibody (AB61791)

ab61791 staining Eph receptor B1 + Eph receptor B2 (phospho Y594) in HeLa cells (overexpressing EphB2 and stimulated for 20 minutes with preclustered efnb1-Fc chimeric protein) by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100 and blocked with 1% BSA for 30 minutes at 25°C. Samples were incubated with primary antibody (1/100 in PBS + 1% BSA + 0.1% Triton X-100) for 2 hour at 25°C. An Alexa Fluor® 488-conjugated donkey anti-rabbit IgG polyclonal (1/300) was used as the secondary antibody. Red - anti-EphB2 (goat), Grey - DAPI, Green - ab61791.

This image is courtesy of an Abreview submitted by Thomas Jungas

• WB

Unknown

Western blot - Anti-Eph receptor B1 + Eph receptor B2 (phospho Y594 + Y596) antibody (AB61791)

All lanes:

Western blot - Anti-Eph receptor B1 + Eph receptor B2 (phospho Y594 + Y596) antibody (ab61791) at 1/500 dilution

Lane 1:

Extracts from HepG2 cells

Lane 2:

Extracts from HepG2 cells with the immunising phosphopeptide

Predicted band size: 110 kDa

Observed band size: 110 kDa

false

• WB

AbReview41567****

Western blot - Anti-Eph receptor B1 + Eph receptor B2 (phospho Y594 + Y596) antibody (AB61791)

All cells were stimulated with preclustered ephrinb1-Fc chimeric protein.

All lanes:

Western blot - Anti-Eph receptor B1 + Eph receptor B2 (phospho Y594 + Y596) antibody (ab61791) at 1/1000 dilution

Lane 1:

HeLa whole cell lysate at 100000 Cells

Lane 2:

HeLa whole cell lysate - overexpressing EphB2 at 100000 Cells

Lane 3:

HeLa whole cell lysate - overexpressinf EphB2-kinase dead (K661R) at 100000 Cells

Secondary

All lanes:

HRP-conjugated goat anti-rabbit IgG monoclonal at 1/5000 dilution

Predicted band size: 110 kDa

Observed band size: 110 kDa

true

Exposure time: 5min

This image is courtesy of an Abreview submitted by Thomas Jungas

• WB

CiteAb

Western blot - Anti-Eph receptor B1 + Eph receptor B2 (phospho Y594 + Y596) antibody (AB61791)

Eph receptor B1 + Eph receptor B2 (phospho Y594 + Y596) western blot using anti-Eph receptor B1 + Eph receptor B2 (phospho Y594 + Y596) antibody ab61791. Publication image and figure legend from Wu, Z., Ashlin, T. G., et al., 2019, Exp Cell Res, PubMed 31075258.

ab61791 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab61791 please see the product overview.

Activation of EphB2 by ephrinB1 and ephrinB1-6F. (A) : Cells expressing EphB2, ephrinB1 or ephrinB1-6F, or control HEK293 cells, were mixed in different combinations in a tube and centrifuged to force them into contact. The cells were lysed at the indicated time points, then subjected to Western blot analysis to detect EphB2, phospho-EphB2 and ephrinB1, and Gapdh as loading control. (B) : Quantitation of phospho-EphB2 detected, normalised to the amount of EphB2 protein. The low amount of endogenous ephrinBs in HEK293 cells elicit low level EphB2 phosphorylation. There is a similar time course of EphB2 phosphorylation after interaction with ephrinB1 or ephrinB1-6F cells, with a >30-fold increase by 5 min that progressively declines at 15 min and 30 min. (C-H) : immunostaining of co-cultures of EphB2/ephrinB1 (C–E) and EphB2/ephrinB1-6F cells (F-H) to detect p-EphB2 and a marker of intracellular vesicles, Rab11. EphB2 cells co-express GFP (green signal in C, F). Activation of EphB2 with ephrinB1 or ephrinB1-6F leads to translocation from the cell surface to intracellular vesicles. Scale bar, 10 μm. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

false