![Simple Western: TRF-2 Antibody (4A794.15) [NB100-56506] - Image shows a specific band for TRF2 in 1.0 mg/mL of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system. Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.](http://images.novusbio.com/fullsize/TRF-2-Antibody-4A794-15-Simple-Western-NB100-56506-img0010.jpg "Simple Western: TRF-2 Antibody (4A794.15) [NB100-56506] - Image shows a specific band for TRF2 in 1.0 mg/mL of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system. Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.")
| Reactivity | Hu, Mu, Rt, Ma, MarSpecies Glossary |
| Applications | WB, Simple Western, ELISA, Flow, Func, ICC/IF, IHC, IP, WB, ChIP, CyTOF-ready |
| Clone | 4A794.15 |
| Clonality | Monoclonal |
| Host | Mouse |
| Conjugate | Unconjugated |
| Format | BSA Free |
| Concentration | 1 mg/ml |
| Description | The TRF2 antibody is referred to as both clone 4A794.15 and 4A794 in the published literature. The TRF2 antibody recognizes full-length TRF2 as well as TRF2 forms lacking both the N-terminal basic domain (B) and the telobox Myb-like C-terminal DNA-binding domain (M). TRF2 forms missing the B and M domains are often referred to as mutant TRF2. Although the exact epitope recognized by the TRF2 antibody has not been mapped, the scientific literature indicates it is in the D or L domain, but not in the B or M domain. |
| Immunogen | This TRF-2 Antibody (4A794.15) was developed against Baculovirus expressed whole length TRF2 protein, used for immunizing mice and splenocytes used to generate the hybridoma clone (NP_005643). |
| Marker | Telomeres marker |
| Specificity | The TRF2 antibody recognizes full-length TRF2 as well as TRF2 forms lacking both the N-terminal basic domain (B) and the telobox Myb-like C-terminal DNA-binding domain (M). TRF2 forms missing the B and M domains are often referred to as mutant TRF2. Although the exact epitope recognized by the TRF2 antibody has not been mapped, the scientific literature indicates it is in the D or L domain, but not in the B or M domain. |
| Isotype | IgG1 Kappa |
| Clonality | Monoclonal |
| Host | Mouse |
| Gene | TERF2 |
| Purity | Protein G purified |
| Innovator's Reward | Test in a species/application not listed above to receive a full credit towards a future purchase. |
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| Application Notes | TRF-2 may be detected as a single band or as a doublet in Western blot. Okabe (2000) described the doublet as 65 and 69 kDa using clone 4A794.15. Observed molecular weights could vary depending on molecular weight standards used and gel conditions. In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue. The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors. |
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| Theoretical MW | 59.6 kDa. Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors. |
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| Storage | Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles. |
| Buffer | PBS |
| Preservative | 0.02% Sodium Azide |
| Concentration | 1 mg/ml |
| Purity | Protein G purified |
| Images | Ratings | Applications | Species | Date | Details | ||||||||
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reviewed by:
Fernanda gutierrez rodrigues |
WB | Human | 09/08/2017 |
Summary
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Secondary Antibodies |
Isotype Controls |
Research Areas for TRF-2 Antibody (NB100-56506)Find related products by research area.
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Application Highlight: Recent uses of TERF2 in immunofluorescence (IF) Telomeres are a region of repeat nucleotide sequences located at the end of chromosomes to protect our DNA from becoming damaged via end-to-end fusion. TERF2, or telomeric-repeat binding factor 2, is important for telomere integrity and aids in th... Read full blog post. |
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TRF2: Telomere Maintenance and Protection TRF2 (telomeric repeat-binding factor 2) is a homodimeric protein that binds the double-stranded 5'-TTAGGG-3' repeat in telomeres. It plays a critical role in the maintenance of telomeres, protecting telomeres against end-to-end chromosomal fusion, an... Read full blog post. |
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| Fernanda gutierrez rodrigues 09/08/2017 |
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| Application: | WB | |
| Species: | Human |
![Western Blot: TRF-2 Antibody (4A794.15) [NB100-56506] - Analysis in human Jurkat cell lysate at 2 ug/mL. Goat anti-mouse Ig HRP secondary antibody and PicoTect ECL substrate solution were used.](http://images.novusbio.com/fullsize/TRF-2-Antibody-4A794-15-Western-Blot-NB100-56506-img0008.jpg "Western Blot: TRF-2 Antibody (4A794.15) [NB100-56506] - Analysis in human Jurkat cell lysate at 2 ug/mL. Goat anti-mouse Ig HRP secondary antibody and PicoTect ECL substrate solution were used.")
![Immunohistochemistry-Paraffin: TRF-2 Antibody (4A794.15) [NB100-56506] - Transitional cell carcinoma, urinary bladder, stained with TRF2 antibody (4 ug/mL), peroxidase-conjugate and DAB chromogen. Note specific nuclear staining. Tumor/normal adjacent tissue array slide was used for this test. Staining of formalin-fixed tissues is enhanced by boiling tissue sections in 10 mM sodium citrate buffer, pH 6.0 for 10-20 min followed by cooling at RT for 20 min.](http://images.novusbio.com/fullsize/TRF-2-Antibody-4A794-15-Immunohistochemistry-Paraffin-NB100-56506-img0012.jpg "Immunohistochemistry-Paraffin: TRF-2 Antibody (4A794.15) [NB100-56506] - Transitional cell carcinoma, urinary bladder, stained with TRF2 antibody (4 ug/mL), peroxidase-conjugate and DAB chromogen. Note specific nuclear staining. Tumor/normal adjacent tissue array slide was used for this test. Staining of formalin-fixed tissues is enhanced by boiling tissue sections in 10 mM sodium citrate buffer, pH 6.0 for 10-20 min followed by cooling at RT for 20 min.")
![Immunocytochemistry/Immunofluorescence: TRF-2 Antibody (4A794.15) [NB100-56506] - Localization of candidate RNAs at telomeres. Localization of SNORD17 at telomeres in human U-2 OS cells. Cells were fixed and sequentially incubated with Abs against TRF-2 and AlexaFluor 488-conjugated anti-mouse IgG. Subsequently, the cells were hybridized with the RNA probes and subjected to fluorescence microscopy. Three different cells are shown. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0123387), licensed under a CC-BY license.](http://images.novusbio.com/fullsize/TRF-2-Antibody-4A794-15-Immunocytochemistry-Immunofluorescence-NB100-56506-img0018.jpg "Immunocytochemistry/Immunofluorescence: TRF-2 Antibody (4A794.15) [NB100-56506] - Localization of candidate RNAs at telomeres. Localization of SNORD17 at telomeres in human U-2 OS cells. Cells were fixed and sequentially incubated with Abs against TRF-2 and AlexaFluor 488-conjugated anti-mouse IgG. Subsequently, the cells were hybridized with the RNA probes and subjected to fluorescence microscopy. Three different cells are shown. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0123387), licensed under a CC-BY license.")
![Flow Cytometry: TRF-2 Antibody (4A794.15) [NB100-56506] - Intracellular flow cytometric analysis of TRF2 in 10^6 human Jurkat cells using 0.1 ug of NB100-56506. The shaded histogram represents cells alone, blue represents isotype control and red represents NB100-56506, anti-TRF2.](http://images.novusbio.com/fullsize/TRF-2-Antibody-4A794-15-Flow-Cytometry-NB100-56506-img0001.jpg "Flow Cytometry: TRF-2 Antibody (4A794.15) [NB100-56506] - Intracellular flow cytometric analysis of TRF2 in 10^6 human Jurkat cells using 0.1 ug of NB100-56506. The shaded histogram represents cells alone, blue represents isotype control and red represents NB100-56506, anti-TRF2.")
![Western Blot: TRF-2 Antibody (4A794.15) [NB100-56506] - Total protein from mouse 3T3 cells was separated on a 12% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2.0 ug/mL anti-TRF2 in 1% non-fat milk in TBST and detected with an anti-mouse HRP secondary antibody using chemiluminescence.](http://images.novusbio.com/fullsize/TRF-2-Antibody-4A794-15-Western-Blot-NB100-56506-img0016.jpg "Western Blot: TRF-2 Antibody (4A794.15) [NB100-56506] - Total protein from mouse 3T3 cells was separated on a 12% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2.0 ug/mL anti-TRF2 in 1% non-fat milk in TBST and detected with an anti-mouse HRP secondary antibody using chemiluminescence.")
![Immunocytochemistry/Immunofluorescence: TRF-2 Antibody (4A794.15) [NB100-56506] - Localization of candidate RNAs at telomeres. Localization of NEAT1 at telomeres in human U-2 OS cells. Cells were fixed and sequentially incubated with Abs against TRF-2 and AlexaFluor 488-conjugated anti-mouse IgG. Subsequently, the cells were hybridized with the RNA probes and subjected to fluorescence microscopy. Three different cells are shown. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0123387), licensed under a CC-BY license.](http://images.novusbio.com/fullsize/TRF-2-Antibody-4A794-15-Immunocytochemistry-Immunofluorescence-NB100-56506-img0019.jpg "Immunocytochemistry/Immunofluorescence: TRF-2 Antibody (4A794.15) [NB100-56506] - Localization of candidate RNAs at telomeres. Localization of NEAT1 at telomeres in human U-2 OS cells. Cells were fixed and sequentially incubated with Abs against TRF-2 and AlexaFluor 488-conjugated anti-mouse IgG. Subsequently, the cells were hybridized with the RNA probes and subjected to fluorescence microscopy. Three different cells are shown. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0123387), licensed under a CC-BY license.")
![Flow Cytometry: TRF-2 Antibody (4A794.15) [NB100-56506] - An intracellular stain was performed on A431 cells with TRF2 Antibody [4A794.15] NB100-56506 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1.0 ug/mL for 30 minutes at room temperature, followed by Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (35503, Thermo Fisher).](http://images.novusbio.com/fullsize/TRF-2-Antibody-4A794-15-Flow-Cytometry-NB100-56506-img0020.jpg "Flow Cytometry: TRF-2 Antibody (4A794.15) [NB100-56506] - An intracellular stain was performed on A431 cells with TRF2 Antibody [4A794.15] NB100-56506 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1.0 ug/mL for 30 minutes at room temperature, followed by Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (35503, Thermo Fisher).")
![Immunocytochemistry/Immunofluorescence: TRF-2 Antibody (4A794.15) [NB100-56506] - Staining of TRF2-bound telomeres in human HeLa cells (Courtesy of Fotiadou, et al, 2004).](http://images.novusbio.com/fullsize/TRF-2-Antibody-4A794-15-Immunocytochemistry-Immunofluorescence-NB100-56506-img0005.jpg "Immunocytochemistry/Immunofluorescence: TRF-2 Antibody (4A794.15) [NB100-56506] - Staining of TRF2-bound telomeres in human HeLa cells (Courtesy of Fotiadou, et al, 2004).")
![Immunocytochemistry/Immunofluorescence: TRF-2 Antibody (4A794.15) [NB100-56506] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.5% Triton-X100. The cells were incubated with anti-TRF-2 (4A794.15) conjugated to FITC [NB100-56506F] at 5 ug/mL for 1 hour at room temperature. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.](http://images.novusbio.com/fullsize/TRF-2-Antibody-4A794-15-Immunocytochemistry-Immunofluorescence-NB100-56506-img0017.jpg "Immunocytochemistry/Immunofluorescence: TRF-2 Antibody (4A794.15) [NB100-56506] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.5% Triton-X100. The cells were incubated with anti-TRF-2 (4A794.15) conjugated to FITC [NB100-56506F] at 5 ug/mL for 1 hour at room temperature. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.")
![Immunocytochemistry/ Immunofluorescence: TRF-2 Antibody (4A794.15) - BSA Free [NB100-56506] - ZBTB48‐dependent loss of MTFP1 phenocopies MTFP1 depletionFluorescence microscopy analysis of the structure & localization of the mitochondrial network in HeLa WT & ZBTB48 KO clones. Mitochondria are marked with the MitoTracker dye (red), & nuclei are counterstained with DAPI (blue). Scale bars represent 20 μm.The same analysis as in (A) for U2OS WT & ZBTB48 KO clones. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28500257), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-56506_mouse-monoclonal-trf-2-antibody-4a794-15-31020241534325.jpg "Immunocytochemistry/ Immunofluorescence: TRF-2 Antibody (4A794.15) - BSA Free [NB100-56506] - ZBTB48‐dependent loss of MTFP1 phenocopies MTFP1 depletionFluorescence microscopy analysis of the structure & localization of the mitochondrial network in HeLa WT & ZBTB48 KO clones. Mitochondria are marked with the MitoTracker dye (red), & nuclei are counterstained with DAPI (blue). Scale bars represent 20 μm.The same analysis as in (A) for U2OS WT & ZBTB48 KO clones. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28500257), licensed under a CC-BY license. Not internally tested by Novus Biologicals.")
![Western Blot: TRF-2 Antibody (4A794.15) - BSA Free [NB100-56506] - TRF2 delta B delta M expression induces chromosome end-to-end fusions in all inducible cell lines(A) Immunoblots of MCF-10A, TO, SH-TO & SV-TO cell lines with & without DOX & HEK 293T. After 96 h of DOX treatment, the inducible cell lines expressed the truncated TRF2 delta B delta M protein (50 kDa); in contrast, uninduced cell lines, MCF-10A parental cell line & HEK 293T cells displayed only the endogen TRF2 protein (66 kDa). Lamin B1 was used as loading control. (B) After sustained expression of TRF2 delta B delta M for 96 h there was a significant increase in aberrant metaphases only in TO cells when compared to uninduced matched cells. (C) Nevertheless, the efficacy of the inducible system was validated by the statistical increase of cells with end-to-end fusions in all inducible cell lines. (D) Moreover, a high incidence of fusions per cell was found in all modified cell lines after sustained TRF2 depletion. Data was presented as mean + SEM. (E) Example of TO, SH-TO & SV-TO karyotypes after 96 h of TRF2 delta B delta M expression. Open arrows indicate clonal aberrations in the parental MCF-10A cell line. Insets in the karyotype show rearranged chromosomes stained with centromeric (green) & telomeric (red) PNA probes. Note the presence of telomere FISH signals at the fusion point of chromatid- or chromosome-type end-to-end fusions. Image collected & cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.25502), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-56506_mouse-monoclonal-trf-2-antibody-4a794-15-31020241682158.jpg "Western Blot: TRF-2 Antibody (4A794.15) - BSA Free [NB100-56506] - TRF2 delta B delta M expression induces chromosome end-to-end fusions in all inducible cell lines(A) Immunoblots of MCF-10A, TO, SH-TO & SV-TO cell lines with & without DOX & HEK 293T. After 96 h of DOX treatment, the inducible cell lines expressed the truncated TRF2 delta B delta M protein (50 kDa); in contrast, uninduced cell lines, MCF-10A parental cell line & HEK 293T cells displayed only the endogen TRF2 protein (66 kDa). Lamin B1 was used as loading control. (B) After sustained expression of TRF2 delta B delta M for 96 h there was a significant increase in aberrant metaphases only in TO cells when compared to uninduced matched cells. (C) Nevertheless, the efficacy of the inducible system was validated by the statistical increase of cells with end-to-end fusions in all inducible cell lines. (D) Moreover, a high incidence of fusions per cell was found in all modified cell lines after sustained TRF2 depletion. Data was presented as mean + SEM. (E) Example of TO, SH-TO & SV-TO karyotypes after 96 h of TRF2 delta B delta M expression. Open arrows indicate clonal aberrations in the parental MCF-10A cell line. Insets in the karyotype show rearranged chromosomes stained with centromeric (green) & telomeric (red) PNA probes. Note the presence of telomere FISH signals at the fusion point of chromatid- or chromosome-type end-to-end fusions. Image collected & cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.25502), licensed under a CC-BY license. Not internally tested by Novus Biologicals.")
![Immunocytochemistry/ Immunofluorescence: TRF-2 Antibody (4A794.15) - BSA Free [NB100-56506] - ZBTB48 ZnF11 necessary to bind to telomeres. Sequence‐specific DNA pull‐downs w/ either telomeric (TTAGGG)/a control sequence (GTGAGT) for FLAG‐ZBTB48 WT, domain deletion constructs for different zinc finger & combinations of deletion constructs w/ ZnF10/11 point mutants. Domain structures indicated - right.Sequence‐specific DNA pull‐downs for FLAG‐ZBTB48 WT & ZnF11 point mutant for telomeric repeat sequences of different phyla (green) & their respective scrambled controls (blue).Protein expression analysis of ZBTB48 by WB for cell lines used. GAPDH serves as a loading control.IF stainings for exogenous FLAG‐ZBTB48 WT & point mutants for ZnF10 & ZnF11 in U2OS cells. The same analysis as in Fig 1E performed & average co‐localization frequencies shown (n = 24–37 cells).Co‐localization analysis of endogenous ZBTB48/exogenous FLAG‐ZBTB48 WT w/ TRF2 in HeLa cells by IF (IF) staining. Image illustrating co‐localization between ZBTB48/FLAG‐ZBTB48 WT (green) & TRF2 (red) as a marker for telomeres is shown w/ DAPI (blue) used as nuclear counterstain. Co‐localization events indicated by white arrows. Quantification of frequency of co‐localization events (right) done after 3D reconstruction of acquired z‐stacks (n = 30 cells).Co‐localization analysis of endogenous ZBTB48/exogenous FLAG‐ZBTB48 WT w/ TRF2 in HeLa 1.3 cells by IF (IF) staining analogous to (E) (n = 30 cells).Co‐localization analysis of endogenous ZBTB48/exogenous FLAG‐ZBTB48 WT w/ TRF2 in HT1080 super‐telomerase cells by IF (IF) staining analogous to (E) (n = 30 cells).Data information: (D–G) Scale bars = 5 μm. Error bars indicate standard deviations, & P‐values based on Student's t‐test.Source data available online for this figure. Image collected & cropped by CiteAb from following publication (//pubmed.ncbi.nlm.nih.gov/28500257), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-56506_mouse-monoclonal-trf-2-antibody-4a794-15-31020241553721.jpg "Immunocytochemistry/ Immunofluorescence: TRF-2 Antibody (4A794.15) - BSA Free [NB100-56506] - ZBTB48 ZnF11 necessary to bind to telomeres. Sequence‐specific DNA pull‐downs w/ either telomeric (TTAGGG)/a control sequence (GTGAGT) for FLAG‐ZBTB48 WT, domain deletion constructs for different zinc finger & combinations of deletion constructs w/ ZnF10/11 point mutants. Domain structures indicated - right.Sequence‐specific DNA pull‐downs for FLAG‐ZBTB48 WT & ZnF11 point mutant for telomeric repeat sequences of different phyla (green) & their respective scrambled controls (blue).Protein expression analysis of ZBTB48 by WB for cell lines used. GAPDH serves as a loading control.IF stainings for exogenous FLAG‐ZBTB48 WT & point mutants for ZnF10 & ZnF11 in U2OS cells. The same analysis as in Fig 1E performed & average co‐localization frequencies shown (n = 24–37 cells).Co‐localization analysis of endogenous ZBTB48/exogenous FLAG‐ZBTB48 WT w/ TRF2 in HeLa cells by IF (IF) staining. Image illustrating co‐localization between ZBTB48/FLAG‐ZBTB48 WT (green) & TRF2 (red) as a marker for telomeres is shown w/ DAPI (blue) used as nuclear counterstain. Co‐localization events indicated by white arrows. Quantification of frequency of co‐localization events (right) done after 3D reconstruction of acquired z‐stacks (n = 30 cells).Co‐localization analysis of endogenous ZBTB48/exogenous FLAG‐ZBTB48 WT w/ TRF2 in HeLa 1.3 cells by IF (IF) staining analogous to (E) (n = 30 cells).Co‐localization analysis of endogenous ZBTB48/exogenous FLAG‐ZBTB48 WT w/ TRF2 in HT1080 super‐telomerase cells by IF (IF) staining analogous to (E) (n = 30 cells).Data information: (D–G) Scale bars = 5 μm. Error bars indicate standard deviations, & P‐values based on Student's t‐test.Source data available online for this figure. Image collected & cropped by CiteAb from following publication (//pubmed.ncbi.nlm.nih.gov/28500257), licensed under a CC-BY license. Not internally tested by Novus Biologicals.")
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![Flow Cytometry: Mouse IgG1 Kappa Isotype Control (P3.6.2.8.1) [NBP1-43319] - Analysis of Alexa Fluor (R) 700 conjugate of NBP1-43319. Human buffy coat stained with mouse IgG1 Kappa isotype control. Flow cytometry image submitted by a verified customer review.](https://images.novusbio.com/images/Mouse-IgG1-Kappa-Light-Chain-Isotype-Control-P3-6-2-8-1-Flow-Cytometry-NBP1-43319-img0002.jpg "Flow Cytometry: Mouse IgG1 Kappa Isotype Control (P3.6.2.8.1) [NBP1-43319] - Analysis of Alexa Fluor (R) 700 conjugate of NBP1-43319. Human buffy coat stained with mouse IgG1 Kappa isotype control. Flow cytometry image submitted by a verified customer review.")
