custom rabbit anti eef1a2 primary antibody  (Proteintech)

Journal: Disease Models & Mechanisms

Article Title: Face-valid phenotypes in a mouse model of the most common mutation in EEF1A2 -related neurodevelopmental disorder

doi: 10.1242/dmm.050501

Figure Lengend Snippet: Eef1a expression analysis. (A,B) Mean Eef1a1 (A) and Eef1a2 (B) mRNA quantities in the brains of P24 mice in the E122K line, normalised to the geometric mean of Gapdh , Ubc and B2m transcript levels. Values for individual biological replicates are shown as dots. Genotypes were compared using ordinary one-way ANOVAs. There was no statistically significant difference in relative Eef1a1 mRNA quantities between genotypes [ F (2, 9)=0.2807, P =0.7616]. There was a statistically significant difference in relative Eef1a2 mRNA quantities between genotypes [ F (2, 9)=16.16, P =0.001]. *** P <0.001 (Dunnett's post-hoc test, +/+ versus other genotypes). (C) Representative Western blots of P24 brain, heart and muscle lysates showing eEF1A2 and a total protein stain. Brain lysates from wst/wst mice, homozygous for an Eef1a2 abolishing deletion , were used as a negative control for eEF1A2. Lysates from adult skeletal muscle were used as a positive control. Brightness and contrast settings were set separately for each blot shown for illustration purposes (applied to the whole image), but this did not affect the quantification using Image Studio Lite software. Western blots were performed in triplicate for each sex with values averaged across replicates. (D-I) Quantification of eEF1A2 levels, normalised to total protein and expressed as a percentage of wild-type expression levels, were compared using ordinary one-way ANOVAs. There were no statistically significant differences in relative eEF1A2 levels in male heart [E; F (2, 6)=1.595, P =0.2782] or female heart [H; F (2, 6)=1.949, P =0.2227]. There were statistically significant differences in relative eEF1A2 levels in male brain [D; F (2, 6)=13.32, P =0.0062], female brain [G; F (2, 6)=39.60, P =0.0003], male muscle [F; F (2, 6)=28.49, P =0.0009] and female muscle [I; F (2, 6)=39.76, P =0.0003]. ** P <0.01, *** P <0.001 (Dunnett's post-hoc test, +/+ versus other genotypes). Sample sizes are shown at the base of the bars. Error bars show the s.d. AU, arbitrary units.

Article Snippet: The custom rabbit anti-eEF1A2 primary antibody was manufactured to order by Proteintech (UK), and was equivalent to antibody eEF1A2-1 described by .

Techniques: Expressing, Western Blot, Staining, Negative Control, Positive Control, Software

Journal: BMC Cancer

Article Title: Translation elongation factor eEF1A2 is a potential oncoprotein that is overexpressed in two-thirds of breast tumours

doi: 10.1186/1471-2407-5-113

Figure Lengend Snippet: Western blot analysis using an anti-eEF1A2 antibody on a range of cell lines. The loading control is GAPDH.

Article Snippet: Primary anti-eEF1A2 rabbit antibodies were used at a concentration of 1:10 diluted in PBS, for 40 minutes at room temperature and secondary goat anti-rabbit IgG biotin conjugated antibody (Dako Cytomation, Cambridgeshire, UK) was used at 1:200 at room temperature for 30 minutes.

Techniques: Western Blot

Journal: BMC Cancer

Article Title: Translation elongation factor eEF1A2 is a potential oncoprotein that is overexpressed in two-thirds of breast tumours

doi: 10.1186/1471-2407-5-113

Figure Lengend Snippet: (A) Real-time RT-PCR analysis of RNA from breast tumours. Each block on the × axis represents a different tumour. The amount of eEF1A2 message is shown normalised to GAPDH and expressed relative to the level of expression in the normal breast RNA samples (=1). ER-negative tumours are shown in white, ER-positive tumours are shown in black. The difference in mean expression between ER-positive and ER-negative samples is 7.2 units (p = 0.0087), 95% Confidence Interval 2.0 to 12.4 units. (B) Average standardised RNA levels in ER-negative and ER-positive breast tumours. This difference is significant (P = 0.0087, t-test).

Article Snippet: Primary anti-eEF1A2 rabbit antibodies were used at a concentration of 1:10 diluted in PBS, for 40 minutes at room temperature and secondary goat anti-rabbit IgG biotin conjugated antibody (Dako Cytomation, Cambridgeshire, UK) was used at 1:200 at room temperature for 30 minutes.

Techniques: Quantitative RT-PCR, Blocking Assay, Expressing

Journal: BMC Cancer

Article Title: Translation elongation factor eEF1A2 is a potential oncoprotein that is overexpressed in two-thirds of breast tumours

doi: 10.1186/1471-2407-5-113

Figure Lengend Snippet: Immunohistochemistry of eEF1A2 in the breast. The panel labelled N shows the antibody staining weakly in a normal breast section. Panels T1 to T5 show breast tumours staining strongly with the anti-eEF1A2 antibody. Magnification ×10.

Article Snippet: Primary anti-eEF1A2 rabbit antibodies were used at a concentration of 1:10 diluted in PBS, for 40 minutes at room temperature and secondary goat anti-rabbit IgG biotin conjugated antibody (Dako Cytomation, Cambridgeshire, UK) was used at 1:200 at room temperature for 30 minutes.

Techniques: Immunohistochemistry, Staining

Journal: BMC Cancer

Article Title: Translation elongation factor eEF1A2 is a potential oncoprotein that is overexpressed in two-thirds of breast tumours

doi: 10.1186/1471-2407-5-113

Figure Lengend Snippet: Western blot analysis using an anti-eEF1A2 antibody on a range of cell lines. The loading control is GAPDH.

Article Snippet: The blots were incubated with primary anti-eEF1A2 rabbit antibody and primary anti-eEF1A1 sheep antibody diluted 1:200 in blocking solution, as well as primary anti-glyceraldehyde-3-phosphate dehydrogenase polyclonal mouse antibody (Chemicon International, Hampshire, UK) diluted 1:10000.

Techniques: Western Blot

Journal: BMC Cancer

Article Title: Translation elongation factor eEF1A2 is a potential oncoprotein that is overexpressed in two-thirds of breast tumours

doi: 10.1186/1471-2407-5-113

Figure Lengend Snippet: (A) Real-time RT-PCR analysis of RNA from breast tumours. Each block on the × axis represents a different tumour. The amount of eEF1A2 message is shown normalised to GAPDH and expressed relative to the level of expression in the normal breast RNA samples (=1). ER-negative tumours are shown in white, ER-positive tumours are shown in black. The difference in mean expression between ER-positive and ER-negative samples is 7.2 units (p = 0.0087), 95% Confidence Interval 2.0 to 12.4 units. (B) Average standardised RNA levels in ER-negative and ER-positive breast tumours. This difference is significant (P = 0.0087, t-test).

Article Snippet: The blots were incubated with primary anti-eEF1A2 rabbit antibody and primary anti-eEF1A1 sheep antibody diluted 1:200 in blocking solution, as well as primary anti-glyceraldehyde-3-phosphate dehydrogenase polyclonal mouse antibody (Chemicon International, Hampshire, UK) diluted 1:10000.

Techniques: Quantitative RT-PCR, Blocking Assay, Expressing

Journal: BMC Cancer

Article Title: Translation elongation factor eEF1A2 is a potential oncoprotein that is overexpressed in two-thirds of breast tumours

doi: 10.1186/1471-2407-5-113

Figure Lengend Snippet: Immunohistochemistry of eEF1A2 in the breast. The panel labelled N shows the antibody staining weakly in a normal breast section. Panels T1 to T5 show breast tumours staining strongly with the anti-eEF1A2 antibody. Magnification ×10.

Article Snippet: The blots were incubated with primary anti-eEF1A2 rabbit antibody and primary anti-eEF1A1 sheep antibody diluted 1:200 in blocking solution, as well as primary anti-glyceraldehyde-3-phosphate dehydrogenase polyclonal mouse antibody (Chemicon International, Hampshire, UK) diluted 1:10000.

Techniques: Immunohistochemistry, Staining