PA5-14607

antibody from Invitrogen Antibodies

Targeting: EPHB2 DRT, EPHT3, ERK, Hek5, Tyro5

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Data presented on provider website)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-14607

Provider

Invitrogen Antibodies

Product name

EphB2 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Reactivity

Human

Host

Rabbit

Isotype

IgG

Vial size

400 µL

Concentration

2 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

References

Submitted references

Expression of ID4 protein in breast cancer cells induces reprogramming of tumour-associated macrophages.

Donzelli S, Milano E, Pruszko M, Sacconi A, Masciarelli S, Iosue I, Melucci E, Gallo E, Terrenato I, Mottolese M, Zylicz M, Zylicz A, Fazi F, Blandino G, Fontemaggi G
Breast cancer research : BCR 2018 Jun 19;20(1):59

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis using an EphB2 polyclonal antibody (Product # PA5-14607) in NCI-H460 cell lysate. Secondary HRP-anti-rabbit was used for signal visualization with chemiluminescence.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of A375 cells using an EphB2 polyclonal antibody (Product # PA5-14607) at a dilution of 1:10-50, followed by a fluor-conjugated goat anti-rabbit secondary antibody (green). Actin filaments were stained with dye-conjugated phalloidin (red). Nuclei were stained with DAPI (blue).

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Fig. 3 Inhibitor of differentiation 4 (ID4) expression in breast cancer cells leads to the activation of an angiogenic programme in macrophages. a Expression matrix representing a panel of angiogenic factors evaluated using TaqMan Low-Density Arrays (TLDA) in macrophages obtained by 1,25-dihydroxyvitamin D 3 (VitD3)-mediated differentiation of HL60 cells and subsequently cultured in RPMI medium or in conditioned media (CM) from control (EV) or ID4-overexpressing (ID4) MDA-MB-468 breast cancer cells. b Western blot showing ID4-HA overexpression in MDA-MB-468 cells. c Selected genes modulated in the arrays were evaluated by RT-qPCR in macrophages obtained from VitD3-mediated differentiation of U937 cells and subsequently cultivated in RPMI medium (CTR) or in CM from control (CM si-SCR) or ID4-depleted (CM si-ID4) MDA-MB-468 cells. d Western blot analysis showing the level of ID4 protein after transfection of the indicated small interfering RNAs (siRNAs) in MDA-MB-468 cells. e - g Western blot analysis of ephrin B2 (EphB2), granulin (GRN) and hypoxia-inducible factor (HIF)-1A proteins in differentiated U937 cells cultured in CM si-SCR or CM si-ID4 from MDA-MB-468 cells. h Immunofluorescence analysis of HIF-1A protein performed in differentiated U937 cells cultured in the presence of CM si-SCR or CM si-ID4 from MDA-MB-468 cells. i Western blotting showing the efficiency of vascular endothelial growth factor A (VEGFA) depletion by siRNA transfection in MDA-MB-468 cells used to pre

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Additional file 7: Figure S4 a Modulation of selected genes modulated in the TLDA was validated by RT-qPCR in differentiated HL60 cells cultured in CM from ID4-overexpressing (CM ID4-HA) or control (CM EV) MDA-MB-468 cells (left panel). The same transcripts were analysed in MDA-MB-468 cells transfected with ID4-HA expression vector (ID4-HA) or control empty vector (EV) (right panel). b Expression of EphB2, MDK and GRN protein evaluated by Western blotting on lysates from differentiated HL60 cells cultured as in ( a ); secreted GRN (sGRN) was evaluated on CM from differentiated HL60 cells in the same conditions. c HIF1A protein expression evaluated by Western blotting in differentiated U937 cells cultured in RPMI medium or in CM from SKBR3 cells stably interfered for ID4 expression (sh-ID4) or control cells (sh-CTR). (PDF 1320 kb)