Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human

Cited:

Human, Mouse, Transgenic Mouse, Xenograft

Applications

Validated:

Immunohistochemistry, Western Blot, Flow Cytometry, CyTOF-ready

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Neutralization, Flow Cytometry, Immunocytochemistry, Immunoprecipitation

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
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Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant human TROP-2
Thr88-Thr274
Accession # P09758

Specificity

Detects human TROP-2 in direct ELISAs and Western blots. In direct ELISAs and Western blots, less than 1% cross-reactivity with recombinant human (rh) MCAM, rhNCAM‑L1, and rhBCAM is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human TROP‑2 Antibody

Detection of Human TROP‑2 antibody by Western Blot.

Detection of Human TROP‑2 by Western Blot.

Western blot shows lysates of NCI-N87 human gastric carcinoma cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human TROP-2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF650) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for TROP-2 at approximately 45-50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.

Detection of TROP-2 antibody in PC-3 Human Cell Line antibody by Flow Cytometry.

Detection of TROP‑2 in PC‑3 Human Cell Line by Flow Cytometry.

PC-3 human prostate cancer cell line was stained with Goat Anti-Human TROP-2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF650, filled histogram) or control antibody (Catalog # AB-108-C, open histogram), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107).

TROP‑2 antibody in Human Brain by Immunohistochemistry (IHC-P).

TROP‑2 in Human Brain.

TROP-2 was detected in immersion fixed paraffin-embedded sections of human brain (frontal cortex) using Goat Anti-Human TROP-2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF650) at 10 µg/mL overnight at 4 °C. Before incubation with the primary antibody tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of Human TROP-2 by Immunohistochemistry

Detection of Human TROP-2 by Immunohistochemistry

Downregulation of Trop2 inhibits cell invasion. (A) The invasive capability of Hep2 cells transfected with NC or Trop2 siRNA (Trop2-S1) was measured at the indicated time points using the Transwell assay. (B) The number of cells on the underside of the chamber was counted. NC, negative control, siRNA, small interfering RNA. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/25779928), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human TROP-2 by Western Blot

Detection of Human TROP-2 by Western Blot

Knockdown of Trop2 expression in Hep2 cells by siRNA. (A) Trop2 mRNA expression in Hep2 cells was examined by reverse transcription-quantitative polymerase chain reaction 48 h after transfection with Trop2 siRNA or NC, as indicated (*P<0.05, **P<0.001). (B) Western blot analysis of Trop2 protein expression in Hep2 cells 48 h following transfection with siRNA as indicated. siRNA, small interfering RNA; NC, negative control. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/25779928), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human TROP-2 by Knockdown Validated

Detection of Human TROP-2 by Knockdown Validated

Trop2 Loss Promotes ErbB3 Activation in HNSCC CellsA. Immunofluorescence images of Trop2 staining (green) in SCC1 HNSCC cells after stable knockdown using short hairpins targeting the Trop2 cDNA. Nuclei are counterstained with 4′6-diamidino-2-phenylindole (DAPI). B. Results of a phosphorylated receptor tyrosine kinase antibody array demonstrating elevated p-ErbB3 in lysates from Trop2 knockdown SCC1 cells. The exposures were normalized to the control spots (four corners), which exhibit equal intensities. C&D. Representative immunoblots showing hyperactivation of ErbB3 and AKT caused by Trop2 loss in SCC1 and SCC25 HNSCC cells. Two short hairpins targeting distinct regions of the Trop2 cDNA were used. E. Reduction of ErbB3 activity after ectopic expression of an RNAi-resistant Trop2 cDNA in Trop2 knockdown SCC1 cells. Arrow points to the lower band which is the correct size for Trop2. F. Ectopic expression of a Flag-epitope tagged Trop2 cDNA in SCC1 cells suppresses basal ErbB3 and AKT activation. Control is an empty vector. Relative increases in phosphoproteins in control versus experimental groups were quantified by photodensitometry after normalization to total ErbB3 or AKT protein which served as an internal controls. Immunoblots are representative of at least three independent experiments. Significance was measured by student's t test, * (P<0.05), ** (P<0.01), *** (P<0.001). Image collected and cropped by CiteAb from the following publication (https://www.oncotarget.com/lookup/doi/10.18632/oncotarget.2423), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human TROP-2 by Western Blot

Detection of Human TROP-2 by Western Blot

Trop2 expression in laryngeal squamous cell carcinoma tissue. (A) Western blot analysis of the protein expression of Trop2 in laryngeal carcinoma tissues (c1, c2, c3 and c4) and paired paracancerous tissues (p1, p2, p3 and p4). Actin was used as a loading control. Representative (B) negative and (C) high expression of Trop2 in paraffin embedding laryngeal carcinoma and precancerous tissues, demonstrated using immunohistochemical staining. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/25779928), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human TROP-2 by Knockdown Validated

Detection of Human TROP-2 by Knockdown Validated

Trop2 Loss Promotes ErbB3 Activation in HNSCC CellsA. Immunofluorescence images of Trop2 staining (green) in SCC1 HNSCC cells after stable knockdown using short hairpins targeting the Trop2 cDNA. Nuclei are counterstained with 4′6-diamidino-2-phenylindole (DAPI). B. Results of a phosphorylated receptor tyrosine kinase antibody array demonstrating elevated p-ErbB3 in lysates from Trop2 knockdown SCC1 cells. The exposures were normalized to the control spots (four corners), which exhibit equal intensities. C&D. Representative immunoblots showing hyperactivation of ErbB3 and AKT caused by Trop2 loss in SCC1 and SCC25 HNSCC cells. Two short hairpins targeting distinct regions of the Trop2 cDNA were used. E. Reduction of ErbB3 activity after ectopic expression of an RNAi-resistant Trop2 cDNA in Trop2 knockdown SCC1 cells. Arrow points to the lower band which is the correct size for Trop2. F. Ectopic expression of a Flag-epitope tagged Trop2 cDNA in SCC1 cells suppresses basal ErbB3 and AKT activation. Control is an empty vector. Relative increases in phosphoproteins in control versus experimental groups were quantified by photodensitometry after normalization to total ErbB3 or AKT protein which served as an internal controls. Immunoblots are representative of at least three independent experiments. Significance was measured by student's t test, * (P<0.05), ** (P<0.01), *** (P<0.001). Image collected and cropped by CiteAb from the following publication (https://www.oncotarget.com/lookup/doi/10.18632/oncotarget.2423), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of TROP-2 by Immunohistochemistry

Detection of TROP-2 by Immunohistochemistry

Differential expression of GFR alpha 1 and SALL4 in As and Apr spermatogonia. Expression of GFR alpha 1 was limited to short chains of Aundiff and more restricted than expression of SALL4. (A–D) Examples of GFR alpha 1 and SALL4 expressing spermatogonial clones in whole mount seminiferous tubules. Scale bars = 10 µm. Panel D was modified from [1] with permission. (E) Quantitative evaluation of GFR alpha 1 and SALL4 expression in clones of undifferentiated spermatogonia. Among As spermatogonia, 50% of cells co-expressed both markers, but substantial populations of SALL4-only and GFR alpha 1-only cells were also observed. The number of SALL4-only cells increased in longer chains, when GFR alpha 1 expression ceased. Asym: Number of clones with molecular asymmetry. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/23326552), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human TROP‑2 Antibody

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

2.5 µg/106 cells
Sample: PC‑3 human prostate cancer cell line

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human brain (cortex) subjected to Antigen Retrieval Reagent-Basic (Catalog # CTS013)

Western Blot

1 µg/mL
Sample: NCI‑N87 human gastric carcinoma cell line

Reviewed Applications

Read 1 review rated 4 using AF650 in the following applications:

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Advanced Features

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  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.


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Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: TROP-2

Human TROP-2, also called tumor associated calcium signal transducer 2 (TACSTD2), GA733-1, gp50 and T16, is a type I cell surface glycoprotein that is highly expressed on human carcinomas. It was originally identified as an antigen present on human gastrointestinal tumors and is the second of two members of this family. The other family member is GA733-2, also called EpCAM, TROP-1, 17-1A, gp40 and KSA. The TROP-2 gene is unique in that it contains no introns. A study of these two genes suggested that TROP-2 was the result of a retroposition of the EpCAM gene. TROP-2 and EpCAM share approximately 49% amino acid identity and approximately 67% similarity. Human and mouse TROP-2 share 87% similarity. The human TROP-2 protein consists of a putative 26 amino acid (aa) signal sequence, a 248 aa extracellular domain, a 23 aa transmembrane region and a 26 aa cytoplasmic domain. TROP-2 is capable of transducing an intracellular calcium signal and may play a role in tumor growth. It also has adhesive functions.

References

  1. Linnenbach, A.J. et al. (1989) Proc. Natl. Acad. Sci. USA 86:27.
  2. Linnenbach, A.J. et al. (1993) Mol. Cell. Biol. 13:1507.
  3. Fornaro, M. et al. (1995) Int. J. Cancer 62:610.
  4. Ripani, E. et al. (1998) Int. J. Cancer 76:671.
  5. El Sewedy, T. et al. (1998) Int. J. Cancer 75:324.

Long Name

Tumor-associated Calcium Signal Transducer 2

Alternate Names

GA733-1, gp50, T16, TACSTD2, TROP2

Entrez Gene IDs

4070 (Human); 56753 (Mouse)

Gene Symbol

TACSTD2

UniProt

Additional TROP-2 Products

Product Documents for Human TROP‑2 Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human TROP‑2 Antibody

For research use only

Citations for Human TROP‑2 Antibody

Customer Reviews for Human TROP‑2 Antibody (1)

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  • Name: Anonymous
    Application: Immunocytochemistry
    Sample Tested: See PMID 23151048
    Species: Human
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