Anti zeb2

Manufactured by Merck Group

Sourced in United States

Anti-ZEB2 is a laboratory reagent designed for the detection and quantification of the ZEB2 protein. It is a specific antibody that binds to the ZEB2 protein, enabling its identification and measurement in biological samples.

Automatically generated - may contain errors

Lab products found in correlation

7 protocols using anti zeb2

Western Blot Analysis of EMT Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols

Immunoblotting was carried out according to standard procedures.12 (link) The following primary antibodies were used in this study: anti-EGFP (ImmunoKontact, UK; catalogue number 210-PS-1GFP), anti-ZEB1 (Santa Cruz Biotechnology, Dallas, USA; sc-25388); anti-SNAIL2 and anti-Cyclin D1 (Cell Signaling Technology, Danvers, USA; 9585 and 2978) anti-ZEB2 (in-house12 (link)); anti-HA and anti-α-Tubulin antibodies from Sigma-Aldrich (St Louis, MO, USA; H3663 and T5168); anti-TWIST1 antibody from Abcam (Cambridge, MA, USA; AB50887); and anti-E-cadherin (BD Bioscience, San Jose, USA; 610181).

Wan Makhtar W.R., Browne G., Karountzos A., Stevens C., Alghamdi Y., Bottrill A.R., Mistry S., Smith E., Bushel M., Pringle J.H., Sayan A.E, & Tulchinsky E. (2017). Short stretches of rare codons regulate translation of the transcription factor ZEB2 in cancer cells. Oncogene, 36(47), 6640-6648.

+ Open protocol

+ Expand

Immunostaining of Stem and Epithelial Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols

The following primary antibodies were used: anti-K14 (polyclonal
rabbit, 1:1000, Thermo Fisher Scientific), anti-GFP (chicken, 1:1000,
Abcam), anti-SOX2 (rabbit, 1:100, Abcam), anti-Vimentin (Rabbit, 1:200,
Abcam), Anti-ECadherin (rat, clone DECMA-1, 1:1000, eBioscience), anti-p63
(polyclonal rabbit, 1:200, Santa Cruz), Anti-Zeb1 (polyclonal rabbit, 1:300,
Bethyl), Anti-Zeb2 (polyclonal rabbit, 1:200, Sigma), anti-EpCam (rabbit
polyclonal, 1:200, Abcam). The following secondary antibodies were used:
anti-rabbit, anti-rat, anti-chicken, conjugated to AlexaFluor488 (1:400,
Molecular Probes), to rhodamine Red-X or to Cy5 (1:400, Jackson
ImmunoResearch).

Latil M., Nassar D., Beck B., Boumahdi S., Wang L., Brisebarre A., Dubois C., Nkusi E., Lenglez S., Checinska A., Drubbel A.V., Devos M., Declercq W., Yi R, & Blanpain C. (2016). Cell-Type-Specific Chromatin States Differentially Prime Squamous Cell Carcinoma Tumor-Initiating Cells for Epithelial to Mesenchymal Transition. Cell stem cell, 20(2), 191-204.e5.

+ Open protocol

+ Expand

Western Blot Analysis of EMT Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were washed twice with phosphate‐buffered saline (PBS) containing CaCl₂ and then lysed in a 100 mM NaCl, 1% NP40, 0.1% SDS, 50 mM Tris pH 8.0 RIPA buffer supplemented with a complete protease inhibitor cocktail (Roche, Mannheim, Germany) and phosphatase inhibitors (Sigma‐Aldrich). Protein expression was examined by Western blot using the anti‐ZEB1 (H102, 1/200, Santa Cruz), anti‐ZEB2 (1/500, Sigma), anti‐TWIST1 (Twist2C1a, 1/100, Abcam, Cambridge, MA, USA), anti‐P‐ERK1/2 (#9106, 1/2,000, Cell Signaling Technology, Danvers, MA, USA), anti‐MITF (clone C5, ab80651, 1/500, Abcam), anti‐FRA1 (sc‐605, 1/400, Santa Cruz), anti‐p75 (1/200, Alomone Labs, Jerusalem), anti‐AXL (AF154, 1/1,000, R&D Systems), and anti‐PARP (cleaved form, 29 kDa) (ab6079, 1/200, Abcam) antibodies for primary detection. Loading was controlled using the anti‐β‐actin (clone AC‐15, 1/10,000, Sigma‐Aldrich), anti‐GAPDH (1/20,000, Millipore), or anti‐α‐tubulin (T5168, 1/5,000, Sigma‐Aldrich) antibodies. Horseradish peroxidase‐conjugated rabbit anti‐mouse, goat anti‐rabbit, and donkey anti‐goat polyclonal antibodies (Dako, Glostrup, Denmark) were used as secondary antibodies. Western blot detections were conducted using the Luminol reagent (Santa Cruz). For ZEB1 level analyses in mouse tumors, proteins were extracted from frozen tumors in liquid nitrogen by homogenizing tissue in RIPA buffer.

Richard G., Dalle S., Monet M., Ligier M., Boespflug A., Pommier R.M., de la Fouchardière A., Perier‐Muzet M., Depaepe L., Barnault R., Tondeur G., Ansieau S., Thomas E., Bertolotto C., Ballotti R., Mourah S., Battistella M., Lebbé C., Thomas L., Puisieux A, & Caramel J. (2016). ZEB1‐mediated melanoma cell plasticity enhances resistance to MAPK inhibitors. EMBO Molecular Medicine, 8(10), 1143-1161.

+ Open protocol

+ Expand

Immunoblotting Antibody Validation Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

Immunoblotting was carried out according to standard procedures.^{12 (link)} The following primary antibodies were used in this study: anti-EGFP (ImmunoKontact, UK; catalog number 210-PS-1GFP), anti-ZEB1 (Santa Cruz Biotechnology, Dallas, TX, USA; sc-25388); anti-SNAIL2 and anti-Cyclin D1 (Cell Signaling Technology, Danvers, USA; 9585 and 2978) anti-ZEB2 (in-house^{12 (link)}); anti-HA and anti-α-Tubulin antibodies from Sigma-Aldrich (St Louis, MO, USA; H3663 and T5168); anti-TWIST1 antibody from Abcam (Cambridge, MA, USA; AB50887); and anti-E-cadherin (BD Bioscience, San Jose, USA; 610181).

+ Open protocol

+ Expand

Immunofluorescence Staining of MDA-MB-231 Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

MDA-MB-231 cells plated on a coverslip were fixed in 4% paraformaldehyde, followed by a rinse with 1× PBS prior to permeabilization in 1× PBS with 0.3% triton X-100. Cells were incubated in blocking buffer (1× PBS with 0.3% triton X-100 and 5% BSA) and then incubated overnight in primary antibodies (anti-ZEB2, Sigma-Aldrich and anti-Fibrillarin, Abcam) diluted in 1× PBS with 0.3% triton X-100 and 1% BSA at 4 °C. The next day, cells were washed in 1× PBS, then incubated in secondary antibodies (Alexa Fluor 594 goat anti-rabbit and Alexa Fluor 488 goat anti-mouse, Thermo Fisher) diluted in blocking buffer. After washing in 1× PBS, the coverslip was mounted using VECTASHIELD PLUS Antifade Mounting Medium with DAPI (Vector Laboratories). Nikon Eclipse Ti-U microscope was used to visualize the cells.

Metge B.J., Alsheikh H.A., Kammerud S.C., Chen D., Das D., Nebane N.M., Bostwick J.R., Shevde L.A, & Samant R.S. (2024). Targeting EMT using low-dose Teniposide by downregulating ZEB2-driven activation of RNA polymerase I in breast cancer. Cell Death & Disease, 15(5), 322.

+ Open protocol

+ Expand

Comprehensive Protein Expression Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Protein was isolated by SDS-PAGE gel and transferred to PVDF membrane. After 1 h of blocking in 5% skim milk, the membrane was incubated with various primary antibodies and incubated overnight at 4 °C with Anti-Alix (sc-53540, Santa Cruz), Anti-α-SMA (ab7817, Abcam), Anti-CD63 (ab134045, Abcam), Anti-FAP (ab207178, Abcam), Anti-ICAM1 (ab282575, Abcam), Anti-MMP-9 (ab76003, Abcam), Anti-MMP-3 (ab52915, Abcam), Anti-β-catenin (ab32572, Abcam), Anti-Ki67 (ab15580, Abcam), Anti-PCNA (ab92552, Abcam), Anti-MMP-2 (ab92536, Abcam), Anti-PDGFR beta (ab69506, Abcam), Anti‐β‐actin (A5441, Sigma), Anti-E-cadherin (SAB4503751, Sigma), Anti-N-cadherin (C3865, Sigma), Anti-Vimentin (V6389, Sigma), Anti-snail (SAB1306281, Sigma), Anti-Zeb1 (SAB2500097, Sigma), Anti-Zeb2 (SAB3500515, Sigma), Anti-Slug (PRS3959, Sigma), Anti‐HSP70 (4873, Cell Signaling Technology), Anti‐GM130 (G7295, Sigma), Anti-FUS (DF8391, Affinity), Anti-ACO1 (AF4711, Affinity), Anti-YTHDC1 (77422, Cell Signaling Technology), Anti-TCEAL7 (DF9969, Affinity). And after three times of elution with TBST, the secondary antibody linked with horseradish peroxidase was incubated for 1 h and the electrochemical luminescence was observed by chemiluminescence apparatus. Results quantization was performed by ImageJ software.

Yan Z., Sheng Z., Zheng Y., Feng R., Xiao Q., Shi L., Li H., Yin C., Luo H., Hao C., Wang W, & Zhang B. (2021). Cancer-associated fibroblast-derived exosomal miR-18b promotes breast cancer invasion and metastasis by regulating TCEAL7. Cell Death & Disease, 12(12), 1120.

+ Open protocol

+ Expand

Comprehensive Protein Expression Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Protein was isolated by SDS-PAGE gel and transferred to PVDF membrane. After 1 h of blocking in 5% skim milk, the membrane was incubated with various primary antibodies and incubated overnight at 4℃ with Anti-Alix (1: 1000, Santa Cruz), Anti-α-SMA (1: 1000, Abcam), Anti-CD63 (1:2000, Abcam), Anti-FAP (1: 1000, Abcam), Anti-ICAM1 (1: 1000, Abcam), Anti-MMP9 (1: 1000, Abcam), Anti-PDGFR beta(1: 1000, Abcam), Anti-β-actin(1: 1000, Sigma), Anti-E-cadherin (1: 1000, Sigma), Anti-N-cadherin(1: 1000, Sigma), Anti-Vimentin(1: 1000, Sigma), Anti-snail (1: 1000, Sigma), Anti-Zeb1(1: 1000, Sigma), Anti-Zeb2(1: 1000, Sigma), Anti-Slug(1: 1000, Sigma), Anti-HSP70 (1: 1000,Cell Signaling Technology), Anti-GM130(1: 1000, Sigma), Anti-ZRANB2(1: 1000, A nity), Anti-ACO1(1: 1000, A nity), Anti-MBNL1(1: 1000, A nity), Anti-TCEAL7(1: 1000, A nity). And after three times of elution with TBST the secondary antibody linked with horseradish peroxidase was incubated for 1 hour and the electrochemical luminescence was observed by chemiluminescence apparatus. Results WB quantization was performed by ImageJ software.

Yan Z., Sheng Z., Zheng Y., Feng R., Shi L., Li H., Yin C., Xiao Q., Luo H., & Zhang B. (2021). Cancer-Associated Fibroblasts Exosomal miR-106a Promotes Breast Cancer Invasion and Metastasis by Down -regulation of TCEAL7.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!