Goat Anti-Chicken IgY H&L (HRP) (ab6877)

Representative Western blot analysis for the selected panel of differentially expressed targets in the various T cell subsets.

Western blot analyses were performed by serially probing sets of 3 membranes per donor. The expression levels of GDIR1, GSTO1, LIMS1 and PROF1 were analyzed blot 1 (upper panel), GDIR2, PRDX2 and THIO on blot 2 (middle panel) and SODM on blot 3 (lower level) of the respective membrane set.

The relative expression levels were defined by densitometric quantification normalized to the corresponding beta-actin signals, co-detected on each membrane.

Aliquots of 15 µg total lysate per sample/lane were loaded onto Tris-Tricine gels (16%T/2.5%C) and subsequently separated. Lysates generated form untreated CD45RA⁺ and CD45RO⁺ T cells were throughout the set of membranes loaded onto the lanes 1 and 3 respectively, whereas lysates generated from the corresponding T cell subsets exposure for 3 h to 5 µM H₂O₂ were loaded onto the lanes 2 and 4 as indicated.

The protein entry names of the targets (left) and their corresponding molecular weight (right) are listed.

ab6877 was used at dilution 1/1000 with the primary antibody ab25912 in WB. See the review on ab25912.

IHC image of beta Actin staining in human normal colon formalin fixed paraffin embedded tissue section*.

The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH 6) for 30 mins. The section was incubated with ab13822, 10 µg/ml overnight at +4°C. An HRP-conjugated secondary (ab6877, 1/500 dilution) was used for 1 hr at room temperature. The section was counterstained with hematoxylin and mounted with DPX.

The inset negative control image is taken from an identical assay without primary antibody.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

ab6877 was used at dilution 1/200 with the primary antibody ab17517 in IHC-P. See the review on ab17517.