37-1600

antibody from Invitrogen Antibodies

Targeting: EPHA4 Hek8, TYRO1

Western blot

ELISA

Immunoprecipitation

Immunohistochemistry

Antibody data

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Validation data

EPHA4 protein structure - 37-1600 shown in red.

Supportive validation

Supportive validation

Supportive validation

Supportive validation

Supportive validation

Supportive validation

Supportive validation

Supportive validation

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: EphA4 phosphorylation in primary spinal cord motor neurons (A and B) Representative Western blot images of pEphA4 and total EphA4 (after immuno-precipitation, IP). (B) graphs showing an average ratio of pEphA4 to total EphA4 in cultures of primary spinal cord motor neurons treated with DMSO, Fc (2.5 mug/mL), ephrinA1-Fc (eA1-Fc, 2.5 mug/mL), 123C4, 150E7, 150E8 (1 and 10 muM) and 150D4 (10 muM) for 15 min. Each panel has its own controls and error bars indicate SEM (each experiment was repeated 3-4 times). Statistical analysis was performed using one-way ANOVA followed by Bonferroni's post-hoc analysis (*p < 0.05, **p < 0.01, compared to DMSO).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 1 EphA4 levels are reduced in the spinal cord of SMNDelta7 mice, without affecting SMN protein levels. Representative images (A) and quantifications (B,C) of a Western blot analysis of EphA4 and SMN levels in spinal cord lysates of Ctrl (smn +/+ and smn +/- ) and SMA (smn -/- ) pups are shown. Beta-actin protein levels were used as a loading control (two-way ANOVA with Sidak's multiple comparison test, n = 4-5 mice/group). * p

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 3 EphrinB3 binds and activates EphA4, EphB1 and EphB6 receptors in SC. a , c , d Orthogonal views of SC activation with clustered EphrinB3 shows that EphrinB3 binds EphB6 ( a ), EphA4 ( c ) and EphB1 ( d ) receptors on SC. b Western blot confirming the presence of these receptors in SC. e , f Bound EphrinB3 activation of EphA4 and EphB receptors viewed by immunodetection of phosphorylated forms. g , h Western blot illustrating that SC incubation with myelin increased the phosphorylation of EphA4 ( g ) and EphB1 + B2 ( h ) which was not the case when myelin was previously blocked by anti-EphrinB3 (O.D. p-EphB1 + B2/GAPDH: one-way ANOVA p = 0.003, F(2,15) = 8.57, followed by a Tukey's multiple comparison test) and O.D. of p-EphA4/GAPDH: one-way ANOVA p = 0.0036, F(2,15) = 8.44, followed by a Tukey's multiple comparison test). Data are expressed as ratio of the optical density (O.D.) of the bands (mean values +- SD) from three independent experiments, control ( n = 6), myelin ( n = 6), myelin + anti-EphrinB3 ( n = 6). Scale bar 20 um

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 1 Reduction of EphA4 protein and mRNA levels in the hippocampus and cortex of APPPS1 mice. Representative images ( a ) and quantification ( b ) of a Western blot analysis of hippocampal EphA4 protein levels in Ctrl and EphA4-KO mice (upper blot) and AD and AD;EphA4-KO mice (lower blot) with GAPDH protein levels as a loading control (unpaired t test and Mann-Whitney test respectively, n = 4-8 mice/group). Representative images ( c ) and quantification ( d ) of a Western blot analysis of cortical EphA4 protein levels in Ctrl and EphA4-KO mice (upper blot) and AD and AD;EphA4-KO mice (lower blot). GAPDH protein levels were assessed to control for equal loading (unpaired t test, n = 4 mice/group). e Representative images of RNA scope with specific probes for EphA4 in the DG, CA3, and CA1 regions of the hippocampus of Ctrl and EphA4-KO mice. Hoechst was used to stain cell nuclei. * p

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 1 Neuroprotection in EphA4 -/- mice and human meta-analysis data of EPHA4. a , b Nissl staining of sagittal sections from WT or EphA4 global knockout mice at 14 days post-CCI injury compared to wild type. c - h Retrospective analysis of data archived as NIH GEO datasets from human patients following brain aneurysms. This study evaluated the gene expression on > 41,000 transcripts. Superficial, n = 10; unruptured, n = 5; ruptured, n = 8. i Western blot analysis for EphA4 protein expression in CCI-injured cortical tissue at 2 h compared to sham-injured cortices ( n = 3-5/group). j Representative confocal images of anti-EphA4 immuno-labeling (red; inset) at 24 h post-CCI injury in the ipsilateral cortex of CX3CR1 GFP/+ mice. CX3CR1-expressing microglia and/or infiltrating monocytes/macrophages show high expression of EphA4 in the peri-lesion cortex (J1) compared to adjacent cells in the medial parietal cortex (J2). Scale bar in j = 200 mum; scale bar in (J1) and (J2) = 20 mum. ANOVA with Bonferroni post hoc test. ** p < 0.01, *** p < 0.001 compared to superficial aneurysms

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 3 Bone marrow chimeric EphA4 -/- mice show decreased lesion volume and altered immune profile following CCI injury. a Bone marrow chimeric WT WTBMC and WT KOBMC mice were generated by irradiation and reconstitution with either wild-type or EphA4 knockout BMCs. PCR showing loss of transcript for Epha4 in the whole blood of WT KOBMC mice compared to WT WTBMC . b - d WT KOBMC mice display decreased lesion volumes compared to WT WTBMC at 3 days following CCI injury. e - i Max projection of z -stack confocal images of the injured cortex showing GFP+ BMCS and DAPI (blue) from WT and j - n KO immune cell infiltration. o Non-biased stereological quantification showed reduced total GFP+ numbers and p co-labeled CD68/GFP-positive cells in the ipsilateral cortex of WT KOBMC mice compared to WT WTBMC . q No difference was seen in the number of Ly6G/GFP+ neutrophils. r CD45-positive enriched cell isolation from the ipsilateral cortex at 3 days showed high purity of cx3cr1 mRNA expression compared with GFAP , Neun , Tie2 , and Vecad . s KO BMCs isolated from the injured cortex showed reduced pro-inflammatory CD68 , MCP1 , and Cxcl16 expression concomitant with t increased pro-resolving TGFbeta , Tie2 , Angpt1 , and Angpt2 . * p < 0.05, *** p < 0.001, **** p < 0.0001 compared to the corresponding WT BMCs. e and j scale bar = 500 mum; f - i and k - n scale bar = 50 mum