Anti-PARIS antibody

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PARIS antibody (AB130867)

IHC image of PARIS staining in rat brain formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab130867, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• WB

Unknown

Western blot - Anti-PARIS antibody (AB130867)

Anti-PARIS antibody (ab130867) detects PARIS protein at ~100kDa, consistent with published data (PMID : 21376232). This blot was produced using a 10% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% Milk before being incubated with ab130867 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

All lanes:

Western blot - Anti-PARIS antibody (ab130867) at 1 µg/mL

Lane 1:

Brain (Mouse) Tissue Lysate at 25 µg

Lane 2:

Brain (Mouse) Tissue Lysate at 25 µg with Human PARIS peptide (<a href='/en-us/products/unavailable/human-paris-peptide-ab157686'>ab157686</a>)

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/10000 dilution

Predicted band size: 69 kDa

Observed band size: 100 kDa,102 kDa,80 kDa

true

Exposure time: 4min

• WB

CiteAb

Western blot - Anti-PARIS antibody (AB130867)

PARIS western blot using anti-PARIS antibody ab130867. Publication image and figure legend from Chen, C. C. W., Erlich, A. T., et al., 2018, Skelet Muscle, PubMed 29549884.

ab130867 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 ab130867 please see the product overview.

Effect of training and Parkin on key regulators of mitochondrial biogenesis. a Representative Western blots of PGC-1α, PARIS and p-ACC expression in untrained and trained skeletal muscle of Parkin KO and WT animals. b A graphical representation of whole muscle PGC-1α (n = 4, *p < 0.05, vs untrained WT). c Quantification of whole muscle PARIS (n = 6, *p < 0.05, main effect of training; #p < 0.05, vs untrained WT). d A graphical representation of whole muscle phosphorylation of ACC (n = 6, *P < 0.05, main effect of training; #P < 0.05, vs untrained KO). Values are means ± SEM. †P < 0.05, interaction effect of training and genotype. α-tubulin was used for muscle loading control. WT, wild type; KO, Parkin knockout; UT, untrained; T, trained; p-ACC, phospho-acetyl-CoA carboxylase; PARIS, Parkin-interacting substrate; PGC-1α, peroxisome proliferator gamma coactivator-1α; A.U., arbitrary units

false