ZEB2 Antibody - BSA Free
Novus Biologicals | Catalog # NBP1-77179
![Western Blot: ZEB2 AntibodyBSA Free [NBP1-77179]](https://resources.rndsystems.com/images/products/ZEB2-Antibody-Western-Blot-NBP1-77179-img0014.jpg "Western Blot: ZEB2 AntibodyBSA Free [NBP1-77179]")
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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.
Scientific Data Images for ZEB2 Antibody - BSA Free
![Western Blot: ZEB2 AntibodyBSA Free [NBP1-77179]](https://resources.rndsystems.com/images/products/ZEB2-Antibody-Western-Blot-NBP1-77179-img0009.jpg "Western Blot: ZEB2 AntibodyBSA Free [NBP1-77179]")
Western Blot: ZEB2 AntibodyBSA Free [NBP1-77179]
Western Blot: ZEB2 Antibody [NBP1-77179] - Analysis in EL4 cell lysate with antibody at 1 ug/mL in (A) the absence and (B) the presence of blocking peptide.
Immunocytochemistry/ Immunofluorescence: ZEB2 Antibody - BSA Free [NBP1-77179] -
Immunocytochemistry/ Immunofluorescence: ZEB2 Antibody - BSA Free [NBP1-77179] - Immunofluorescence of ZEB2 in Jurkat cells with ZEB2 antibody at 20 ug/ml.Red: ZEB2 Antibody
Blue: DAPI staining
![Western Blot: ZEB2 AntibodyBSA Free [NBP1-77179]](https://resources.rndsystems.com/images/products/ZEB2-Antibody-Western-Blot-NBP1-77179-img0012.jpg "Western Blot: ZEB2 AntibodyBSA Free [NBP1-77179]")
Western Blot: ZEB2 AntibodyBSA Free [NBP1-77179]
ZEB2-Antibody-Western-Blot-NBP1-77179-img0012.jpg![Western Blot: ZEB2 AntibodyBSA Free [NBP1-77179]](https://resources.rndsystems.com/images/products/ZEB2-Antibody-Western-Blot-NBP1-77179-img0013.jpg "Western Blot: ZEB2 AntibodyBSA Free [NBP1-77179]")
Western Blot: ZEB2 AntibodyBSA Free [NBP1-77179]
ZEB2-Antibody-Western-Blot-NBP1-77179-img0013.jpg
Western Blot: ZEB2 Antibody - BSA Free [NBP1-77179] -
Western Blot: ZEB2 Antibody - BSA Free [NBP1-77179] - Fbxo45 induces degradation of core EMT-TFsA. Immunoblot for core EMT-TFs in 293T cells transfected Zeb1, Zeb2, Snai1, Snai2 or Twist1 with increasing amount of Fbxo45. B. Immunoblot for detecting endogenous core EMT-TFs in HeLa cells transfected with or without myc-tagged Fbxo45, & harvested after treatment with 20μM of MG132 for 4 hours. C. Immunoblot for endogenous Zeb1, Zeb2, Snai1 & Snai2 in MCF7 cells treated with indicated different concentrations of 17 beta -estrogen for 24 h in the phenol-free medium to induce the expression of endogenous Fbxo45. D. Immunoblot for endogenous core EMT-TFs in HeLa transfected with siRNAs for Fbxo45 or Pam. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/25460509), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Western Blot: ZEB2 Antibody - BSA Free [NBP1-77179] -
Western Blot: ZEB2 Antibody - BSA Free [NBP1-77179] - Fbxo45 induces degradation of core EMT-TFsA. Immunoblot for core EMT-TFs in 293T cells transfected Zeb1, Zeb2, Snai1, Snai2 or Twist1 with increasing amount of Fbxo45. B. Immunoblot for detecting endogenous core EMT-TFs in HeLa cells transfected with or without myc-tagged Fbxo45, & harvested after treatment with 20μM of MG132 for 4 hours. C. Immunoblot for endogenous Zeb1, Zeb2, Snai1 & Snai2 in MCF7 cells treated with indicated different concentrations of 17 beta -estrogen for 24 h in the phenol-free medium to induce the expression of endogenous Fbxo45. D. Immunoblot for endogenous core EMT-TFs in HeLa transfected with siRNAs for Fbxo45 or Pam. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/25460509), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Western Blot: ZEB2 Antibody - BSA Free [NBP1-77179] -
Western Blot: ZEB2 Antibody - BSA Free [NBP1-77179] - Fbxo45 induces degradation of core EMT-TFsA. Immunoblot for core EMT-TFs in 293T cells transfected Zeb1, Zeb2, Snai1, Snai2 or Twist1 with increasing amount of Fbxo45. B. Immunoblot for detecting endogenous core EMT-TFs in HeLa cells transfected with or without myc-tagged Fbxo45, & harvested after treatment with 20μM of MG132 for 4 hours. C. Immunoblot for endogenous Zeb1, Zeb2, Snai1 & Snai2 in MCF7 cells treated with indicated different concentrations of 17 beta -estrogen for 24 h in the phenol-free medium to induce the expression of endogenous Fbxo45. D. Immunoblot for endogenous core EMT-TFs in HeLa transfected with siRNAs for Fbxo45 or Pam. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/25460509), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Western Blot: ZEB2 Antibody - BSA Free [NBP1-77179] -
Western Blot: ZEB2 Antibody - BSA Free [NBP1-77179] - Fbxo45 induces degradation of core EMT-TFsA. Immunoblot for core EMT-TFs in 293T cells transfected Zeb1, Zeb2, Snai1, Snai2 or Twist1 with increasing amount of Fbxo45. B. Immunoblot for detecting endogenous core EMT-TFs in HeLa cells transfected with or without myc-tagged Fbxo45, & harvested after treatment with 20μM of MG132 for 4 hours. C. Immunoblot for endogenous Zeb1, Zeb2, Snai1 & Snai2 in MCF7 cells treated with indicated different concentrations of 17 beta -estrogen for 24 h in the phenol-free medium to induce the expression of endogenous Fbxo45. D. Immunoblot for endogenous core EMT-TFs in HeLa transfected with siRNAs for Fbxo45 or Pam. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/25460509), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Western Blot: ZEB2 Antibody - BSA Free [NBP1-77179] -
Western Blot: ZEB2 Antibody - BSA Free [NBP1-77179] - Functional domains of Fbxo45 for ubiquitination of Zeb2A. Lysates from 293T cells transfected Flag-Zeb2 with or without HA-Fbxo45 were immunoprecipitated (IP) with anti-Flag M2 antibody, & then immunoblotted. Total lysates were also used as Input for immunoblotting analysis. B. Bacterially expressed GST or GST–Fbxo45, -Fbxo45 delta F-box, -Fbxo45 delta mid, or -Fbxo45 delta SPRY fusion proteins & Glutathione-Sepharose beads were incubated with lysates of HeLa cells transfected with Zeb2, Snai1 or Snai2, respectively. The proteins associated with GST–tagged Fbxo45 forms, bound on the Glutathione-Sepharose beads were washed five times with the RIPA buffer before immunoblotting. C. Lysates from 293T cells transfected Flag-Zeb2 with HA-Fbxo45, -Fbxo45 delta mid, -Fbxo45 delta F-box or -Fbxo45 delta SPRY were immunoprecipitated with anti-Flag or anti-HA antibodies, & immunoprecipitates were resolved by SDS-PAGE for Western-blot analysis. D. U2OS cells transfected with Flag-Zeb2 & HA-Fbxo45, -Fbxo45 delta F-box or -Fbxo45 delta SPRY were stained using the primary antibodies of anti-Flag M2 or anti-HA, & the second antibodies of Alexa Fluor 568 anti-mouse or Alexa Fluor 488 anti-Rabbit, respectively. Scale: 25μm. E-I. Zeb2 ubiquitination assays by using IP expriments under different conditions: Flag-Zeb2 with or without HA-Ub (E); Flag-Zeb2 with HA-Ub WT, K48R or K63R (F); Flag-Zeb2 & HA-Ub K48-only with or without myc-Fbox45 (G); Flag-Zeb2 & HA-Ub K48-only with siRNA control, siRNAs for Fbxo45 or siRNAs for Pam (H); Flag-Zeb2 & HA-Ub with HA-Fbxo45, -Fbxo45 delta F-box or -Fbxo45 delta SPRY (I). Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/25460509), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Western Blot: ZEB2 Antibody - BSA Free [NBP1-77179] -
Western Blot: ZEB2 Antibody - BSA Free [NBP1-77179] - Fbxo45 is a direct target of miR-27a* that mediates EMT processesA, C. The expression levels of miR-27a* (A) & Fbxo45 mRNA (C) in P69 & M12 were shown according to the data of miRNA transcriptome sequencing & DGE sequencing, respectively [46]. TPM means transcripts per million. B. Real-time PCR in delta delta Ct methods were used to confirm the expression levels of miR-27a* in P69 & M12. One representative experiment out of three was shown. D. Fbxo45 mRNA levels in stable cell lines miR-27a*-overexpressing P69 & miR-27a*-silencing M12 (zipmiR-27a*). P69 or M12 stably infected with the same lentiviral empty vector was used as a control (ctrl). E. Fbxo45 3′-UTR wide-type or mutant form at the position where is complementary to the 5′ seed region of miR-27a* was subcloned into psiCheck2 vector. 293T cells were transfected with psiCheck2 containing WT or mutant Fbxo45 3′-UTR & miR-27a* or non-specific control vector. The Renilla luciferase activity was normalized on the constitutive activity of firefly luciferase. Data are the mean±S.E.M. of three independent experiments. F-G. Immunoblot for Fbxo45, core EMT-TFs (Zeb1, Zeb2, Twist1) & their associated EMT markers (Fibronectin, N-cadherin, Laminin C2, Vimentin & E-cadherin) in stable cell lines P69-miR-27a* or M12-zipmiR-27a* (F); Immunoblot for detection of protein stability of core EMT-TFs under the treatment with 20μM of MG132 for 4 h before harvested (G). GAPDH is for a loading control. H. Immunblot for Fbxo45, N-cadherin, E-cadherin & five core EMT-TFs in stable cell lines PC3-ctrl, PC3-miR-27a* & PC3-zipmiR-27a*. alpha -tubulin is for a loading control. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/25460509), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Western Blot: ZEB2 Antibody - BSA Free [NBP1-77179] -
Western Blot: ZEB2 Antibody - BSA Free [NBP1-77179] - Fbxo45 is a direct target of miR-27a* that mediates EMT processesA, C. The expression levels of miR-27a* (A) & Fbxo45 mRNA (C) in P69 & M12 were shown according to the data of miRNA transcriptome sequencing & DGE sequencing, respectively [46]. TPM means transcripts per million. B. Real-time PCR in delta delta Ct methods were used to confirm the expression levels of miR-27a* in P69 & M12. One representative experiment out of three was shown. D. Fbxo45 mRNA levels in stable cell lines miR-27a*-overexpressing P69 & miR-27a*-silencing M12 (zipmiR-27a*). P69 or M12 stably infected with the same lentiviral empty vector was used as a control (ctrl). E. Fbxo45 3′-UTR wide-type or mutant form at the position where is complementary to the 5′ seed region of miR-27a* was subcloned into psiCheck2 vector. 293T cells were transfected with psiCheck2 containing WT or mutant Fbxo45 3′-UTR & miR-27a* or non-specific control vector. The Renilla luciferase activity was normalized on the constitutive activity of firefly luciferase. Data are the mean±S.E.M. of three independent experiments. F-G. Immunoblot for Fbxo45, core EMT-TFs (Zeb1, Zeb2, Twist1) & their associated EMT markers (Fibronectin, N-cadherin, Laminin C2, Vimentin & E-cadherin) in stable cell lines P69-miR-27a* or M12-zipmiR-27a* (F); Immunoblot for detection of protein stability of core EMT-TFs under the treatment with 20μM of MG132 for 4 h before harvested (G). GAPDH is for a loading control. H. Immunblot for Fbxo45, N-cadherin, E-cadherin & five core EMT-TFs in stable cell lines PC3-ctrl, PC3-miR-27a* & PC3-zipmiR-27a*. alpha -tubulin is for a loading control. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/25460509), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Immunocytochemistry/ Immunofluorescence: ZEB2 Antibody - BSA Free [NBP1-77179] -
Immunocytochemistry/ Immunofluorescence: ZEB2 Antibody - BSA Free [NBP1-77179] - Immunocytochemistry of ZEB2 in Jurkat cells with ZEB2 antibody at 20 u/mL.
Immunocytochemistry/ Immunofluorescence: ZEB2 Antibody - BSA Free [NBP1-77179] -
Immunocytochemistry/ Immunofluorescence: ZEB2 Antibody - BSA Free [NBP1-77179] - Immunofluorescence of ZEB2 in Jurkat cells with ZEB2 antibody at 20 u/mL.Applications for ZEB2 Antibody - BSA Free
ELISA
Immunocytochemistry/ Immunofluorescence
Western Blot
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Background: ZEB2/SIP1
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UniProt
Additional ZEB2/SIP1 Products
Product Documents for ZEB2 Antibody - BSA Free
Certificate of Analysis
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Product Specific Notices for ZEB2 Antibody - BSA Free
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Appropriate Fixation of IHC/ICC Samples
- Cellular Response to Hypoxia Protocols
- ClariTSA™ Fluorophore Kits
- Detection & Visualization of Antibody Binding
- ELISA Sample Preparation & Collection Guide
- ELISA Troubleshooting Guide
- How to Run an R&D Systems DuoSet ELISA
- How to Run an R&D Systems Quantikine ELISA
- How to Run an R&D Systems Quantikine™ QuicKit™ ELISA
- ICC Cell Smear Protocol for Suspension Cells
- ICC Immunocytochemistry Protocol Videos
- ICC for Adherent Cells
- Immunocytochemistry (ICC) Protocol
- Immunocytochemistry Troubleshooting
- Immunofluorescence of Organoids Embedded in Cultrex Basement Membrane Extract
- Immunohistochemistry (IHC) and Immunocytochemistry (ICC) Protocols
- Preparing Samples for IHC/ICC Experiments
- Preventing Non-Specific Staining (Non-Specific Binding)
- Primary Antibody Selection & Optimization
- Protocol for VisUCyte™ HRP Polymer Detection Reagent
- Protocol for the Fluorescent ICC Staining of Cell Smears - Graphic
- Protocol for the Fluorescent ICC Staining of Cultured Cells on Coverslips - Graphic
- Protocol for the Preparation and Fluorescent ICC Staining of Cells on Coverslips
- Protocol for the Preparation and Fluorescent ICC Staining of Non-adherent Cells
- Protocol for the Preparation and Fluorescent ICC Staining of Stem Cells on Coverslips
- Protocol for the Preparation of a Cell Smear for Non-adherent Cell ICC - Graphic
- Quantikine HS ELISA Kit Assay Principle, Alkaline Phosphatase
- Quantikine HS ELISA Kit Principle, Streptavidin-HRP Polymer
- R&D Systems Quality Control Western Blot Protocol
- Sandwich ELISA (Colorimetric) – Biotin/Streptavidin Detection Protocol
- Sandwich ELISA (Colorimetric) – Direct Detection Protocol
- TUNEL and Active Caspase-3 Detection by IHC/ICC Protocol
- The Importance of IHC/ICC Controls
- Troubleshooting Guide: ELISA
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
FAQs for ZEB2 Antibody - BSA Free
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Q:
As the person that originally gave the name of ZEB2 to ZEB2 (Proc Natl Acad Sci U S A. 97:6391-6), I would kindly ask you to withdraw the sale of your Ab BP1-7717. This is NOT ZEB2 ( in its many names: zfhx1b, SIP1, etc.) In your link you indicated ZEB2 as a 95-100 kDa protein, when the predicted is 140 kDa and with all the posttranslational modifications is close to 180 or more. I happened to review an article for a scientific journal that was using your Ab and showing ZEB2 as a 100 kDa. Of course, I rejected the article as the Ab is clearly something else. Problem is that other referees may be unaware of this and your Ab is creating a lot of confusion with articles being published for who knows what protein (but not ZEB2).
A: The immunogen for catalog number NBP1-77179 is from ZEB2 (NCBI accession number NP_055610, specific sequence DDSSEDGKMETKSDHEED). Attached is an image that shows bands at 180kDa (although strong ones are seen at ~90kD - perhaps through a cleavage, splicing or degradation event). I hope this will help you reduce some of the concern associated with this product and the published paper. I apologize for the misrepresented image loaded on the website. It is certainly incorrect and will be adjusted immediately. Lane A and B are both Jurkat, A was probed with the antibody at 1ug/ml and B was at a concentration 2ug/ml.
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Q: I'm seeing multiple bands in WB for # NBP1-77179. I would like to know why.
A: Thank you for contacting Novus regarding the NBP1-77179. The faint band about 90kDa may be a cleavage, splicing or degradation event. There are often bands that we cannot explain due to the epitope and splicing or cleavage. It is typical to see this pattern in many of westerns of different antibodies. I am afraid we have not done analysis and the literature I have examined does not have an easy explanation other than an alternate splice.
-
Q:
As the person that originally gave the name of ZEB2 to ZEB2 (Proc Natl Acad Sci U S A. 97:6391-6), I would kindly ask you to withdraw the sale of your Ab BP1-7717. This is NOT ZEB2 ( in its many names: zfhx1b, SIP1, etc.) In your link you indicated ZEB2 as a 95-100 kDa protein, when the predicted is 140 kDa and with all the posttranslational modifications is close to 180 or more. I happened to review an article for a scientific journal that was using your Ab and showing ZEB2 as a 100 kDa. Of course, I rejected the article as the Ab is clearly something else. Problem is that other referees may be unaware of this and your Ab is creating a lot of confusion with articles being published for who knows what protein (but not ZEB2).
A: The immunogen for catalog number NBP1-77179 is from ZEB2 (NCBI accession number NP_055610, specific sequence DDSSEDGKMETKSDHEED). Attached is an image that shows bands at 180kDa (although strong ones are seen at ~90kD - perhaps through a cleavage, splicing or degradation event). I hope this will help you reduce some of the concern associated with this product and the published paper. I apologize for the misrepresented image loaded on the website. It is certainly incorrect and will be adjusted immediately. Lane A and B are both Jurkat, A was probed with the antibody at 1ug/ml and B was at a concentration 2ug/ml.
-
Q: I'm seeing multiple bands in WB for # NBP1-77179. I would like to know why.
A: Thank you for contacting Novus regarding the NBP1-77179. The faint band about 90kDa may be a cleavage, splicing or degradation event. There are often bands that we cannot explain due to the epitope and splicing or cleavage. It is typical to see this pattern in many of westerns of different antibodies. I am afraid we have not done analysis and the literature I have examined does not have an easy explanation other than an alternate splice.