Anti-Cardiac Troponin I antibody [EP1106Y]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cardiac Troponin I antibody [EP1106Y] (AB52862)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human cardiac muscle tissue sections labeling Cardiac Troponin I with Purified ab52862 at 1 : 300 dilution (1.19 µg/ml). Heat mediated antigen retrieval was performed Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cardiac Troponin I antibody [EP1106Y] (AB52862)

IHC image of Cardiac Troponin I staining in human heart formalin fixed paraffin embedded tissue section, performed on a Leica Bond 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 ab52862, 1/200 dilution, 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

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cardiac Troponin I antibody [EP1106Y] (AB52862)

ab52862 at 1/250 dilution staining cardiac Troponin I in human cardiac muscle by Immunohistochemistry, Paraffin embedded tissue.

• WB

Lab

Western blot - Anti-Cardiac Troponin I antibody [EP1106Y] (AB52862)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Cardiac Troponin I antibody [EP1106Y] (ab52862) at 1/500000 dilution

All lanes:

Human fetal heart lysates at 15 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 24 kDa

Observed band size: 28 kDa

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• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Cardiac Troponin I antibody [EP1106Y] (AB52862)

Intracellular Flow Cytometry analysis of A-673 (Human muscle Ewing's Sarcoma) cells labeling Cardiac Troponin I (red) with ab52862 at a 1/30 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluorr^® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (Black) (ab172730). Blue (unlabeled control) - Cell without incubation with primary antibody and secondary antibody (Blue).

• IP

Unknown

Immunoprecipitation - Anti-Cardiac Troponin I antibody [EP1106Y] (AB52862)

ab52862 (purified) at 1 : 30 dilution (2μg) immunoprecipitating Cardiac Troponin I in Human fetal heart lysate.
Lane 1 (input) : Human fetal heart lysate 10μg
Lane 2 (+) : ab52862 & Human fetal heart lysate
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab52862 in Human fetal heart lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1 : 1000 dilution.
Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-Cardiac Troponin I antibody [EP1106Y] (ab52862)

Predicted band size: 24 kDa

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• WB

Unknown

Western blot - Anti-Cardiac Troponin I antibody [EP1106Y] (AB52862)

All lanes:

Western blot - Anti-Cardiac Troponin I antibody [EP1106Y] (ab52862) at 1/1000000 dilution

All lanes:

fetal heart cell lysate at 10 µg

Secondary

All lanes:

Goat anti-rabbit HRP at 1/2000 dilution

Predicted band size: 24 kDa

Observed band size: 28 kDa

false

• WB

CiteAb

Western blot - Anti-Cardiac Troponin I antibody [EP1106Y] (AB52862)

Cardiac Troponin I western blot using anti-Cardiac Troponin I antibody [EP1106Y] ab52862. Publication image and figure legend from Lewinska, A., Adamczyk-Grochala, J., et al., 2020, Aging (Albany NY), PubMed 32584786.

ab52862 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 ab52862 please see the product overview.

Remifentanil preconditioning-mediated effects on metabolic activity (A), cell number (B), the levels of necrotic cells (C), troponin I release, a marker of myocardial damage (D) and the levels of apoptotic cells (E, F) during normoxic and hypoxic conditions in human cardiac myocytes (HCM). (A) The metabolic activity was assayed using MTT test. Metabolic activity at control normoxic conditions (CTR) is considered as 100%. A solvent action (0.9% NaCl) is also shown. Based on MTT results, the concentration of 8 ng/ml remifentanil was selected for further analysis. Cell number (B) and necrotic cell death (C) were evaluated using TC10™ automated cell counter. (C) Necrosis was analyzed using trypan blue exclusion assay. (D) Western blot analysis of the levels of cardiac troponin I. Cardiac troponin I levels in supernatants (Myocyte Growth Medium, MGM) were calculated per 10000 cells. Two biomarkers of apoptotic cell death were considered, namely phosphatidylserine externalization (E) and the activity of caspase 3/7 (F) using Muse® Cell Analyzer and Muse® Annexin V and Dead Cell Assay Kit and Muse® Caspase-3/7 Assay Kit, respectively. Representative dot-plots are also shown. Bars indicate SD, n = 3, ***p < 0.001, **p < 0.01, *p < 0.05 compared to normoxic control (CTR), ###p < 0.001, #p < 0.05 compared to hypoxic control (CTR) (ANOVA and Dunnett's a posteriori test). CTR, control; R, remifentanil preconditioning.

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