Overview

  • Product name

    Anti-TROP2 antibody [162-46.2]
    See all TROP2 primary antibodies
  • Description

    Mouse monoclonal [162-46.2] to TROP2
  • Host species

    Mouse
  • Tested applications

    Suitable for: IHC-P, ICC/IF, Flow Cytmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Tissue, cells or virus corresponding to Human TROP2. Human choriocarcinoma cell line BeWo.

  • Positive control

    • IHC-P: Human placenta tissue. ICC/IF: JEG3 and MCF7 cells. Flow Cyt: JEG3 and MCF7 cells.
  • General notes

    This antibody clone is manufactured by Abcam.

    Isotype: IgG1 (Igh-4a allotype)

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: PBS, 6.97% L-Arginine
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Monoclonal
  • Clone number

    162-46.2
  • Isotype

    IgG1
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab79976 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
IHC-P Use at an assay dependent concentration.
ICC/IF Use a concentration of 5 µg/ml.
Flow Cyt Use 0.1-1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

Target

  • Function

    May function as a growth factor receptor.
  • Tissue specificity

    Placenta, pancreatic carcinoma cell lines.
  • Involvement in disease

    Defects in TACSTD2 are the cause of gelatinous drop-like corneal dystrophy (GDLD) [MIM:204870]; also known as lattice corneal dystrophy type III. GDLD is an autosomal recessive disorder characterized by grayish corneal amyloid deposits that cause severe visual impairment.
  • Sequence similarities

    Belongs to the EPCAM family.
    Contains 1 thyroglobulin type-1 domain.
  • Post-translational
    modifications

    The N-terminus is blocked.
  • Cellular localization

    Membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • Cell surface glycoprotein Trop 2 antibody
    • Cell surface glycoprotein Trop-2 antibody
    • Cell surface glycoprotein Trop2 antibody
    • Epithelial glycoprotein 1 antibody
    • GA733 1 antibody
    • GA7331 antibody
    • M1S 1 antibody
    • M1S1 antibody
    • Membrane component chromosome 1 surface marker 1 antibody
    • Pancreatic carcinoma marker protein GA733 1 antibody
    • Pancreatic carcinoma marker protein GA733-1 antibody
    • Pancreatic carcinoma marker protein GA7331 antibody
    • TACD 2 antibody
    • TACD2_HUMAN antibody
    • TACSTD 2 antibody
    • Tacstd2 antibody
    • Trop 2 antibody
    • Trop2 antibody
    • Tumor associated calcium signal transducer 2 precursor antibody
    • Tumor-associated calcium signal transducer 2 antibody
    see all

Images

  • Overlay histogram showing JEG3 cells stained with ab79976 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab79976, 0.1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150117) at 1/2000 dilution for 30 min at 22°C.

    Isotype control antibody (black line) was mouse IgG1 [15-6E10A7] (monoclonal) (ab170190) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

    Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter.

    This antibody gave a positive signal in JEG3 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions.

  • MCF7 cells stained for TROP2 (colored green) using ab79976 in ICC/IF. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab79976, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Overlay histogram showing MCF7 cells stained with ab79976 (red line). The cells were fixed with 80% methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab79976, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive result in 4% paraformaldehyde (10 min) fixed MCF7 cells used under the same conditions.

    Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.

  • ab79976 staining TROP2 in human 10 week placenta by Immunohistochemistry, Formalin-fixed, Paraffin-embedded tissue. Anti-mouse peroxidase IgG used as secondary antibody. Note staining of cytotrophoblast (CT) and syncytiotrophoblast (ST).

References

This product has been referenced in:

  • Viswanatha R  et al. Interactome analysis reveals ezrin can adopt multiple conformational states. J Biol Chem 288:35437-51 (2013). ICC/IF ; Human . Read more (PubMed: 24151071) »
  • Lipinski M  et al. Human trophoblast cell-surface antigens defined by monoclonal antibodies. Proc Natl Acad Sci U S A 78:5147-50 (1981). Read more (PubMed: 7029529) »
See all 2 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Answer

Thank you for contacting Abcam.

Thank you for pointing out the error in figure legends for ab824 and ab79976. The flow cytometry figure legends for the two antibodies have been switched on the online datasheet to correspond with the correct antibody.
ab824: (https://www.abcam.com/fgfr1-antibody-m5g10-ab824.html)

ab824 is specifically raised against the ectodomain FGFR1 beta isoform. Thus, the statement in the epitope description that the "epitope is masked in undenatured FGFr1a" refers to the specificity of this antibody for the FGFR1 beta isoform.

Below are some general troubleshooting tips for flow cytometry, if you are experiencing issues with ab824 please reply with further sample, protocol and troubleshooting steps that have been taken. These will not only help me to fully understand the problem but are essential should further testing of this product be needed as well.
https://www.abcam.com/index.html?pageconfig=resource&rid=11443

Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

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Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Pig Tissue sections (Skin)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: EDTA/Tris
Permeabilization
No
Specification
Skin
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Apr 23 2012

Answer

Vielen Dank für Ihre Anrufe.

Wie versprochen habe ich alle unsere Langerin- bzw. TACD2-Antikörper danach geprüft, ob sie schon einmal mit Schwein getestet wurden - was leider bei keinem der Fall ist.

Daher habe ich die Immunogene auf Sequenzähnlichkeit mit dem Langerin bzw. TACD2 des Schweins (SwissProt IDs B3FVQ1 bzw. F1S7A4) untersucht, und die BLAST-Recherchen ergaben folgendes:

Langerin

ab68434 (Immunogen Aminosäuren 1-20, MTVEKEAPDAHFTVDKQNIS): 83%

https://www.abcam.com/index.html?datasheet=68434 (or use the following: https://www.abcam.com/index.html?datasheet=68434).

ab125476: 79%

https://www.abcam.com/index.html?datasheet=125476 (or use the following: https://www.abcam.com/index.html?datasheet=125476).

ab90959: 66%

https://www.abcam.com/index.html?datasheet=90959 (or use the following: https://www.abcam.com/index.html?datasheet=90959).

ab22111: viele Treffer mit unterschiedlichen Ähnlichkeiten (ca. 70%), daher nicht empfehlenswert zu testen

https://www.abcam.com/index.html?datasheet=22111 (or use the following: https://www.abcam.com/index.html?datasheet=22111).

TACD2

ab65006 (Aminosäuren 130-180): 92%

https://www.abcam.com/index.html?datasheet=65006 (or use the following: https://www.abcam.com/index.html?datasheet=65006).

ab58930 (Immunogen Aminosäuren 194-209, VHYEQPTIQIELRQNT): 86%

https://www.abcam.com/index.html?datasheet=58930 (or use the following: https://www.abcam.com/index.html?datasheet=58930).

ab77551: 85%

https://www.abcam.com/index.html?datasheet=77551 (or use the following: https://www.abcam.com/index.html?datasheet=77551).

ab89928: 85%

https://www.abcam.com/index.html?datasheet=89928 (or use the following: https://www.abcam.com/index.html?datasheet=89928).

ab64320: Die Immunogensequenz muss im Labor nachgefragt werden.

https://www.abcam.com/index.html?datasheet=64320 (or use the following: https://www.abcam.com/index.html?datasheet=64320).

Für diese Antikörper kann ich Ihnen wie zuvor wieder jeweils einen Testrabatt anbieten, d.h. wenn Sieeinen der Antikörper kaufen, uns Ihre Ergebnisse mit Schwein per Abreview mitteilen, bekommen Sie wieder jeweils einen kosten Primärantikörper zum Dank (und die Abpoints nicht zu vergessen) - https://www.abcam.com/collaborationdiscount.

Dies wäre vor allem für den TACD2- Antikörper ab65006 zu empfehlen, den wir schon in der Immunhistochemiegetestet haben und daher wissen, dass er wenigstensin der Anwendung funktioniert - auch wenn die Spezies Schwein noch nichtvalidiert wurde. Allerdings ist mit einer Sequenzähnlichkeit von 92%die Wahrscheinlichkeit einer Kreuzreaktivität sehr hoch.Aus dem gleichen Grund würde sich auch ab64320 eignen; hier können wir aber gerne noch die Rückmeldung aus dem Labor abwarten. Alle anderen Antikörper wurden bisher noch nicht in der Immunhistochemie getestet, daher besteht hier ein doppeltes Risiko, da wir nicht wissen, ob die Antikörper für diese Anwendung geeignet sind.

Sie können diese Antikörper auch wie besprochen kostenlos erhalten, wenn Sie sie mittels der Gutscheine von den vorherigen Testungen bestellen. Allerdings können wir in diesem Fall keine Garantie dafür übernehmen, wenn sienicht mit Schwein kreuzreagieren.

Ich hoffe, diese Informationen helfen Ihnen weiter. Falls Sie einen Rabattcode erhalten möchten oder weitere Fragen haben, zögern Sie bitte nicht, sich wieder an mich zu wenden.

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Answer

Glad to hear it! Please let me know if you need any further assistance.

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Question

Here you can find an answer.

Our staining protocol for Tacsd2 is 1:20 (ph6.0). As you can see in the attached file (A, C), we had a beautiful immunoreactivity on cancer nests as well as surrounding bile ducts. However, we didn’t have any immunoreacitivy this time by staining same protocol. We already tried several additional staining conditions (i.e., changing dilution, changing PH, changing tempreature) before contacting you, however, all staining conditions were completely negative to both cancer nests and surrounding bile ducts. We really hope that we can find a solution because this staining is a key part of our next coming paper.

1) Can you please confirm the species of your liver tissue?

a. Human liver tissue fixed by formalin.

2) What antigen retrieval and permeabilization was performed? What blocking agent was used?

a. As antigen retrieval we used a ready-to-use EDTA base pH 9.0 solution for 20 and 40 minutes (HIER). Additionally we did an antigen retrieval with a ready-to-use citrate based pH 6.0 solution. As a blocking agent we used a peroxidase block, which is included in the Bond Polymer Refine detection kit.

3) What were the primary and secondary antibody concentrations and incubation times?

a. The dilution of the primary antibody was 1/20 with an incubation time of 30 minutes. The secondary antibody was a ready-to-use antibody from Leica (Bond Polymer Refine Detection kit) with an incubation of 8 minutes

4) Can you please send an image of your results or describe the staining you are seeing?

a. We have checked immunoreactivity on human cholangiocellular carcinoma. As you can see in the attached file, Tacstd2 immunoreacitivy was seen on cancer nests as well as surrounding normal bile ducst (A). Figure B was a picture at this time showing negative for Tacstd2. We use intrahepatic bile ducts as an internal positive control (C). again this time, internal control was all negative (D). * A and C were the previous staining we had done in last year. B and D were the staining which we did this time.

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Answer

Thank you for sending the data as well as all the protocol description and for explaining the experiments.

Could you please let us know if you have tried both lots (GR41084-1 and GR41084-2) and with the same results of no staining?

We have not received any other complaints for this antibody or these lots - it could maybe just have been a shipment issue.

If all the 5 vials give the same results, then the best solution would be to replace them with another lot. The current lot is GR41084-3, but it is the only one in stock at the moment.

I would advise to try only one vial and see if that works as before. If it does, I'd be happy to send 4 more vials.

Please let me know how you would like to proceed.

I look forward to hear back from you.

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Answer

Thank you for contacting us. I am sorry to hear that the new lot you have received is not producing good results.

After checking our records, we have not had any other complaints about this antibody in recent years. It is possible that there may be an issue with the lot or vials you received. Have you tested all five vials of antibody? Did they all produce the same results?

It would be very helpful if you could please provide some additional details of your protocol:

1.) Can you please confirm the species of your liver tissue?

2.)What antigen retrieval and permeabilization was performed? What blocking agent was used?

3.) What were the primary and secondary antibody concentrations and incubation times?

4.) Can you please send an image of your results or describe the staining you are seeing?

I will be happy to help you further once I have this additional information.

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