Overview

  • Product name
    Anti-Cardiac Troponin I antibody
    See all Cardiac Troponin I primary antibodies
  • Description
    Rabbit polyclonal to Cardiac Troponin I
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, ICC/IF, IHC-Fr, IHC-P, Flow Cyt, ELISAmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Pig
    Predicted to work with: Chimpanzee
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human cardiac Troponin I.

    Read Abcam's proprietary immunogen policy (Peptide available as ab47002.)

  • Positive control
    • This antibody gave a positive signal in the following tissue lysates: Heart (human) tissue lysate, mouse heart tissue.

Properties

Applications

Our Abpromise guarantee covers the use of ab47003 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB Use a concentration of 1 µg/ml. Detects a band of approximately 26 kDa (predicted molecular weight: 24 kDa).Can be blocked with Human Cardiac Troponin I peptide (ab47002).
ICC/IF Use at an assay dependent concentration.
IHC-Fr 1/100.
IHC-P 1/100.
Flow Cyt Use at an assay dependent concentration. PubMed: 21909276

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

ELISA Use at an assay dependent concentration.

Target

  • Function
    Troponin I is the inhibitory subunit of troponin, the thin filament regulatory complex which confers calcium-sensitivity to striated muscle actomyosin ATPase activity.
  • Involvement in disease
    Defects in TNNI3 are the cause of cardiomyopathy familial hypertrophic type 7 (CMH7) [MIM:613690]. Familial hypertrophic cardiomyopathy is a hereditary heart disorder characterized by ventricular hypertrophy, which is usually asymmetric and often involves the interventricular septum. The symptoms include dyspnea, syncope, collapse, palpitations, and chest pain. They can be readily provoked by exercise. The disorder has inter- and intrafamilial variability ranging from benign to malignant forms with high risk of cardiac failure and sudden cardiac death.
    Defects in TNNI3 are the cause of cardiomyopathy familial restrictive type 1 (RCM1) [MIM:115210]. RCM1 is an heart muscle disorder characterized by impaired filling of the ventricles with reduced diastolic volume, in the presence of normal or near normal wall thickness and systolic function.
    Defects in TNNI3 are the cause of cardiomyopathy dilated type 2A (CMD2A) [MIM:611880]. Dilated cardiomyopathy is a disorder characterized by ventricular dilation and impaired systolic function, resulting in congestive heart failure and arrhythmia. Patients are at risk of premature death.
    Defects in TNNI3 are the cause of cardiomyopathy dilated type 1FF (CMD1FF) [MIM:613286]. Dilated cardiomyopathy is a disorder characterized by ventricular dilation and impaired systolic function, resulting in congestive heart failure and arrhythmia. Patients are at risk of premature death.
  • Sequence similarities
    Belongs to the troponin I family.
  • Information by UniProt
  • Database links
  • Alternative names
    • cardiac muscle antibody
    • Cardiac troponin I antibody
    • cardiomyopathy, dilated 2A (autosomal recessive) antibody
    • Cardiomyopathy, familial hypertrophic, 7, included antibody
    • CMD1FF antibody
    • CMD2A antibody
    • CMH7 antibody
    • cTnI antibody
    • Familial hypertrophic cardiomyopathy 7 antibody
    • MGC116817 antibody
    • RCM1 antibody
    • Tn1 antibody
    • Tni antibody
    • TNN I3 antibody
    • TNNC 1 antibody
    • TNNC1 antibody
    • TNNI3 antibody
    • TNNI3_HUMAN antibody
    • Troponin I antibody
    • Troponin I cardiac antibody
    • Troponin I cardiac muscle antibody
    • Troponin I cardiac muscle isoform antibody
    • Troponin I type 3 cardiac antibody
    • troponin I, cardiac 3 antibody
    • TroponinI antibody
    • Ttroponin I type 3 (cardiac) antibody
    see all

Images

  • All lanes : Anti-Cardiac Troponin I antibody (ab47003) at 1 µg/ml

    Lane 1 : Human heart tissue lysate - total protein (ab29431)
    Lane 2 : Human liver tissue lysate - total protein (ab29889)

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 24 kDa
    Observed band size: 26 kDa
    why is the actual band size different from the predicted?

  • IHC image of Cardiac Troponin I staining in human heart formalin fixed paraffin embedded tissue section, performed on a Leica BondTM 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 ab47003, 1µg/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.
  • ab74003 staining Cardiac Troponin I in neonatal Rat cardiomyocytes by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized in 0.1% Triton X-100 and blocked with 3% BSA for 1 hour at 25°C. Samples were incubated with primary antibody (1/400 in 3% BSA + PBS-T) for 12 hours at 4°C. An AlexaFluor® 594-conjugated Goat anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody.

    See Abreview

  • All lanes : Anti-Cardiac Troponin I antibody (ab47003) at 1 µg/ml

    Lane 1 : Human heart tissue lysate - total protein (ab29431)
    Lane 2 : Heart (Mouse) Tissue Lysate
    Lane 3 : Heart (Rat) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 24 kDa
    Observed band size: 26 kDa why is the actual band size different from the predicted?
    Additional bands at: 160 kDa, 80 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 3 minutes
  • All lanes : Anti-Cardiac Troponin I antibody (ab47003) at 1/20000 dilution

    All lanes : Pig Heart whole cell lysate

    Lysates/proteins at 4 µg per lane.

    Secondary
    All lanes : An HRP-conjugated Goat anti-rabbit polyclonal at 1/20000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 24 kDa
    Observed band size: 29 kDa why is the actual band size different from the predicted?



    Blocking Step: 20% Serum for 3 hours at 24°C

    See Abreview

  • Immunohistochemical analysis of frozen mouse heart tissue labeling Cardiac Troponin I with ab47003 at 1/100 dilution.

    See Abreview

  • ab47003 staining Cardiac Troponin I in Human cardiomyocytes by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100 and blocked with 1% BSA for 30 minutes at 20°C. Samples were incubated with primary antibody (1/200 in PBS + 1% BSA) for 1 hour at 20°C. An undiluted FITC-conjugated Goat anti-rabbit IgG polyclonal was used as the secondary antibody.

    See Abreview

References

This product has been referenced in:
  • Wang Z  et al. Imaging transparent intact cardiac tissue with single-cell resolution. Biomed Opt Express 9:423-436 (2018). Read more (PubMed: 29552383) »
  • Schneider C  et al. The Anti-Cancer Multikinase Inhibitor Sorafenib Impairs Cardiac Contractility by Reducing Phospholamban Phosphorylation and Sarcoplasmic Calcium Transients. Sci Rep 8:5295 (2018). Read more (PubMed: 29593308) »
See all 53 Publications for this product

Customer reviews and Q&As

1-10 of 19 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
ELISA
Sample
Human Serum (immobilization on beads)
Specification
immobilization on beads
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Type
Sandwich (Capture)

Mr. Chang Su Jeon

Verified customer

Submitted Sep 19 2012

Application
Immunocytochemistry
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 22°C
Sample
Human Cultured Cells (Fetal Cardiomyocytes)
Specification
Fetal Cardiomyocytes
Permeabilization
Yes - 0.1% Triton
Fixative
Paraformaldehyde

Jennifer Dewing

Verified customer

Submitted Mar 24 2015

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 15 minute(s) · Concentration: 5% · Temperature: RT°C
Sample
Human Cell (Human pluripotent stem cell derived cardiomyocyte)
Specification
Human pluripotent stem cell derived cardiomyocyte
Permeabilization
Yes - saponin
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Nov 19 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (heart)
Specification
heart
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate
Permeabilization
No
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Mar 26 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Pig Tissue sections (heart)
Specification
heart
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate
Permeabilization
No
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Mar 22 2013

Question

Dear Tech Support Team,
Please read below and see the attached images (the troponin is green, cd31is pink) and advise to interpret the results.
Thanks in advance for your assistance.
Antibody code: ab47003,24590
Batch number:
General Information:
Antibody storage conditions (temperature/reconstitution etc) as stated in the date sheet. -20 in diluents
Description of the problem (high background, low signal, non-specific staining etc.)
non-specific staining and the staining itself is not like in the pictures in abcam webpage
Sample (Species/Tissue/Cell Type/Cell Line etc.) cells (cardiomyocytes and HUVEC on a scaffold
Fixation of sample (Ethanol/Methanol/Acetone/Paraformaldehyde/Other/Duration etc.) Methanol 10 min
Antigen retrieval (Enzymatic method, Heat mediated technique etc.) non needed
Permeabilization step tween 20 0.05% in PBS
Blocking conditions (Buffer/time period, Blocking agent etc.) super block-scytek 8 min
Primary Antibody (Diluent/Dilution/Incubation time, Wash step) ab47003 1:200,24590 1:100 in pbs+1%BSA for 1 hr RT,wash in tween 20 0.05% in PBS 3 times for 5 min while shaking
Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)
goat anti-rabbit Alexa Fluor 488 (Jackson Immuno Research Laboratories, Inc.), Goat Anti-Mouse Alexa Fluor 594 (Jackson Immuno Research Laboratories, Inc.), both 1:500
Detection method confocal
Positive and negative controls used (please specify) non, but secondary antibodies were in exact prior use with great specificity, both abcam antibodies were stained not together to generate good results
Optimization attempts (problem solving):none
How many times have you tried the IHC? twice
Have you run a "No Primary" control?
secondary antibodies were in exact prior use with great specificity
Do you obtain the same results every time? some pictures you don’t see much cross reactivity, some you see

Read More
Answer

Thank you for contacting us.
I appreciate the time taken to submit protocol information. The details you have kindly provided will provide us with vital information for our monitoring of product quality. I would also appreciate if you could please provide the order and batch details.
Reviewing this case, I would like to offer some suggestions to help optimise the results from these two antibodies in ICC/IF:
Cell fixation should be done under cold conditions. Using ice cold methanol, and optimizing it with other fixatives, such as 3-4% paraformaldehyde in PBS pH 7.4 for 15 min at room temperature.
For antibody ab24590 permeabilization is not required as the target protein is expressed in the membrane. For ab47003 I would recommend increasing the permeabilization conditions, by using more concentrated Tween 20 or 0.25% Triton X-100.
The unspecific signal obtained may be due to unspecific binding of the antibody as there has not been any blocking of unspecific binding sites. This could be improved by using 3%BSA for 30 min / 1 hour at room temperature.
Also, in order to facilitate the antibody to reach the target, incubation overnight at 4C normally gives better results than 1 hour incubation.
I hope this information is helpful. Should the suggestions not improve the results, please do not hesitate to contact me again and I will try to provide further help.

Read More

Answer

Thank you for getting back to me with the new results obtained.

It seems from the image shared that the staining is much improved with the changes in the protocol. The nuclear staining is much improved and the striations observed would be expected in the staining of this protein. In order to ascertain if there is any non-specific staining I would suggest the following:

1. Carry out a "no primary" control. This would involve staining the cells with the secondary antibody only at the same dilution as used before (1/40), then measure the fluorescence using the same parameters used for the images shared with me, i.e. using the rhodamine filters. This will allow you to see exactly how much of the non-specific staining is derived from the secondary antibody binding to the cells non-specifically. Ifnon-specific binding of the secondary antibody is observed Iwould suggest reducing the concentration of secondary antibody used to 1/200 and 1/500 to see if the results improve.

2. What buffer is being used to dilute theprimary antibody? I would suggest using 3% BSA in PBST.

3. Do you know what staining you would expect to get in human mesenchymal stem cells? Do you have any references where similar staining has been performed? Do you have access to a good positive control such as cardiomyocytes?

I hope these suggestions will be of help.


I look forward to receiving your reply.

Read More

Answer

Thank you for taking time to complete our questionnaire and sorry for the delay in getting back to you. I am sorry to hear that this antibody is not providing satisfactory results. The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality.

We would not expect the kind of nuclear staining observed in your experiments. Having reviewed this case, I would like to offer some suggestions to help optimize the results from ab47003 and obtain the staining expected.

1. I would suggest altering the blocking used to 3% BSA instead of the serum. If you want to use serum then it should be from the species in which the secondary antibody was raised, in this case Donkey. BSA has been successfully used by several of our customers, one of which is illustrated from the following link:

https://www.abcam.com/index.html?datasheet=47003&tab=abreviews&intabreviewid=30342

In the blocking buffer I would also introduce 0.3 M glycine. This will bind any free aldehyde groups that could otherwise bind the primary or secondary antibody potentially leading toa high background.

2. I would also suggest reducing the concentration of the primary antibody used, from 1/100 to 1/400.

3. I would suggest checking the non-specific binding of the secondary antibody. This can be performed bu incubating the slide with the secondary antibody following the normal blocking step and viewing the slide as normal using the Rhodamine filter. If this is contributing to the non-specificity I would suggest reducing the concentration from 1/50.

Should the suggestions not improve the results, please do let me know.

In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund.

I hope this information is helpful, and I thank you for your cooperation.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Adult heart)
Specification
Adult heart
Fixative
Formaldehyde
Permeabilization
Yes - 0.5% Tx100 in PBS
Blocking step
Serum as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 1% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Jun 18 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (neonatal cardiomyocyte)
Specification
neonatal cardiomyocyte
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% TritonX-100
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Mrs. Jamie Soto

Verified customer

Submitted Jun 15 2012

1-10 of 19 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Sign up