Anti-Cardiac Troponin T antibody [1C11]

• ICC

Lab

Immunocytochemistry - Anti-Cardiac Troponin T antibody [1C11] (AB8295)

Immunofluorescence staining of Cardiac Troponin T using ab8295 ab277612, which were differentiated for 10 days post induction.

The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% PBS-Tween for 5 mins and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab8295 at 5 µg/mL and ab6046, rabbit polyclonal to beta Tubulin, at 1/1000 dilution. Cells were then incubated with ab150117, Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150088, Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) preadsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown. Gamma is adjusted to 1.5 in all channels.

The antibody ab8295 gave comparable results using MeOH fixation (100%, 5 min).

• IHC-P

PubMed

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cardiac Troponin T antibody [1C11] (AB8295)

Paraffin-embedded Normal human Heart and iVSD heart tissue (were blocked using 10% FBS for 30 min) stained for Cardiac Troponin T (Red) using ab8295 at 1/200 dilution at room temperature for 2 hours. The slides were then incubated with Fluor^® 555-conjugated anti-mouse (Abcam, ab150107; 1 : 1,000 dilution). The nuclear counterstain was DAPI (Blue).

Ye L et al. Decreased Yes-Associated Protein-1 (YAP1) Expression in Pediatric Hearts with Ventricular Septal Defects. PLoS One 10:e0139712 (2015).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cardiac Troponin T antibody [1C11] (AB8295)

ab8295 staining human normal heart. Staining is localised to the cytoplasm.
Left panel : with primary antibody at 1 ug/ml. Right panel : isotype control.
Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffer citrate pH 6.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS), then incubated with primary antibody for 20 minutes, and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.

• sELISA

Unknown

Sandwich ELISA - Anti-Cardiac Troponin T antibody [1C11] (AB8295)

Calibration curves for sandwich cTnT fluoroimmunoassay with different animal TnTs as antigen.(dark blue) canine, (blue/grey) human, (grey) mouse, (black) rat. Monoclonal antibodies : capture, ab8295 [clone 1C11], 1 μg/well, detection ab1454 [clone 7E7], 200 ng/well. Assay time, 30 min at room temperature.

• ICC/IF

CiteAb

Immunocytochemistry/ Immunofluorescence - Anti-Cardiac Troponin T antibody [1C11] (AB8295)

Immunocytochemistry-immunofluorescence using Anti-Cardiac Troponin T antibody [1C11], ab8295. Publication image from Li, J. et al., 2020, Theranostics, 32724455. Legend direct from paper.

Ablation of CD4+ but not CD8+ T-cells reactivates neonatal heart regeneration after cryoinfarction. (A) Schematic diagram showing the experimental design. (B) Images of scar tissues, scale bars : 2000 µm; and Masson's trichrome staining showing representative serial cross sections of fibrotic tissues in blue, scale bars : 1000 µm. (C) Quantification of fibrotic tissue coverage based on (B). Immunostaining on frozen sections for (D) COLA1+ (red) and cTnT+ (green) cells within the infarct zone, scale bars : 100 µm; or (F) pH3+ (red) or Ki67+ (red) and cTnT+ (green) cells within the border zone, scale bars : 50 µm. (F) Arrows indicate cardiomyocytes positive for pH3 or Ki67 and squares denote magnified images on the right. Quantification of absolute number of (E) %cTnT+ coverage, (G) pH3+cTnT+ or (H) Ki67+cTnT+ cardiomyocytes per mm2 area. (C, E, G, H) Data are presented as mean±S.E.M., *P<0.05, **P<0.01, n=4-5 per group.

• IHC

CiteAb

Immunohistochemistry - Anti-Cardiac Troponin T antibody [1C11] (AB8295)

Immunohistochemistry-immunofluorescence using Anti-Cardiac Troponin T antibody [1C11], ab8295. Publication image from Foo, R. S. et al., 2017, Nat Commun, 28790305. Legend direct from paper.

Quadrant analyses reveal sub-populations of CM. a–c Quadrant analysis for Proliferation vs. Negative regulators of proliferation genes identifies increased co-expression in individual TAC nuclei (Q2; 44.4%; p = 3.237e−07 Fisher’s exact test), only detectable by single nuclear RNA-seq (a), and not in pooled nuclei (b) or matched bulk left ventricle RNA-seq (c). Inset : histogram of nuclei distributed across quadrants. Blue represents Sham and red represents TAC nuclei. d–f Quadrant analysis for cardiac progenitor vs. cardiac transcription factor gene expression shows increased co-expression upon TAC stress in single CM nuclei d (Q2; 42.9%; p = 2.548e−05 Fisher’s exact test), again not detectable in pooled nuclei or bulk tissue RNA-seq (e, f). g–i Increased co-expression of fetal reprogramming genes and dedifferentiation markers under TAC stress only detected in single nuclear RNA-seq (g) (Q2; 58.73%; p = 0.001371 Fisher’s exact test) and not in non-single approaches (h, i). j High co-expression of cardiac progenitors, cardiac transcription factors, dedifferentiation, proliferation, and negative proliferation markers is confined to single nuclear TAC samples in Q2 and Q4. k, l Single molecule RNA FISH shows Sca1 upregulation and co-expression of Tnnt2 in isolated adult mouse CMs from TAC hearts l compared to Sham k. Number of Sca1+ Sham CMs : 5/13; Sca1+ TAC CMs : 38/55; all together from 2 Sham and 3 TAC biological replicates. m, n Immunofluorescence confirms increase in cell-surface SCA1 protein expression in TAC CMs (n) compared to Sham (m). Number of SCA1+ Sham CMs : 8/23; SCA1+ TAC CMs : 43/66; all together from 2 Sham and 3 TAC biological replicates. Scale bar represents 20 µm