Home Primary Antibodies Anti-ZEB1 Rabbit Antibody [M1M18] For research use only.

Anti-ZEB1 Rabbit Antibody [M1M18]

Catalog No.: F0266

    Application: Reactivity:
    • F0266-wb
      Lane 1: Jurkat
      Lane 2: HT1080
      Lane 3: A172
      Lane 4: COS (mock transfected)
      Lane 5: COS (transfected with hZEB1)

    Experiment Essentials

    To ensure effective detection, it is necessary to extract nuclear proteins before conducting experiments.

    WB
    Recommended SDS-PAGE separating gel concentration: 5%.

    Usage Information

    Dilution
    1:1000
    1:200
    Application
    WB, IP
    Reactivity
    Human
    Source
    Rabbit
    Storage Buffer
    PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN₃
    Storage (from the date of receipt)
    –20°C (avoid freeze-thaw cycles), 2 years
    Predicted MW
    200 kDa
    Positive Control COS (transfected with hZEB1); Jurkat; HT1080; A172
    Negative Control COS

    Expression & Treatment Conditions

    Cell Line Expression Level References Sample Treatment
    COS No Expression
    Jurkat Low Expression
    HT-1080 Low Expression
    A-172 Low Expression
    HCT116 Low Expression 39307879
    SW480 Low Expression 39307879
    BT-549 Low Expression 37915591 BT-549 TNBC cells growing in 2D culture were established as serial passage 1 (S1) mammospheres. These mammosphere cells were then isolated and serially passaged for up to S3 mammospheres
    Colo320-HSR Medium Expression 37771778
    Colo320-DM Medium Expression 37771778
    NCIH716 Medium Expression 37771778
    SW620 No Expression 37771778
    SNUC2A No Expression 37771778
    SNUC4 No Expression 37771778
    DLD-1 No Expression 37771778
    HCT8 No Expression 37771778
    KM12C No Expression 37771778
    NCIH498 No Expression 37771778
    PANC1 High Expression 37771778
    MIAPaCa2 High Expression 37771778
    SNU410 Medium Expression 37771778
    SNU324 No Expression 37771778
    SNU213 No Expression 37771778
    Capan2 No Expression 37771778
    Capan1 No Expression 37771778
    AsPC1 Medium Expression 37771778
    BXPC3 Medium Expression 37377917
    U87 High Expression 37062068
    U251 High Expression 37062068
    MDA-MB-231 Medium Expression 37122033
    LN18 Medium Expression 37004060
    PN12 Medium Expression 37004060
    LY1 Medium Expression 36725845
    LY3 Medium Expression 36725845
    NCH421k High Expression 36765531
    U87 High Expression 36765531
    MDA-MB-231 Medium Expression 36604594
    human breast CSCs Medium Expression 36538894
    HCT116 Medium Expression 36346211
    SW480 Medium Expression 36346211
    HCT116 Medium Expression 36362141 treatment with bufalin (20 nM) for 24 h
    DLD-1 Medium Expression 36362141 treatment with bufalin (20 or 40 nM) for 48 h
    SW480 Medium Expression 36362141 treatment with bufalin (20 or 40 nM) for 48 h
    Y79 Medium Expression 36291907 topotecan-resistant or carboplatin-resistant Y79 cells
    MDA-MB-231-BR High Expression 36291142
    SK-OV-3 High Expression 36114703
    A2780 Medium Expression 36114703
    WM793 Medium Expression 36052492
    1205Lu High Expression 36052492
    SKMEL28 No Expression 35580987 SKMEL28 cells were treated with increasing doses of trametinib (up to 20 nM) until they acquired resistance (R+T cells). Trametinib was removed from R+T in an acute (R−T acute) or sustain (R−T sustain) manner.
    HPMVEC Low Expression 35978929
    A2780 Low Expression 35675036 transwell cocultured with primary cancer-associated fibroblasts (CAFs) for 48 h
    OVCAR3 Low Expression 35675036 transwell cocultured with primary cancer-associated fibroblasts (CAFs) for 48 h
    MCF-7 Medium Expression 35951366 transwell cocultured with primary cancer-associated fibroblasts (CAFs) for 48 h
    LN229 Low Expression 35910786
    P3 Low Expression 35910786
    U-138 MG Medium Expression 35768187
    A-172 Medium Expression 35768187
    H1155 Medium Expression 35838183
    H358 No Expression 35838183
    H441 Low Expression 35838183
    HCC461 Medium Expression 35838183
    A549 Low Expression 35838183
    H157 Low Expression 35838183
    H647 Medium Expression 35838183
    HCC44 Medium Expression 35838183
    H460 Low Expression 35838183
    ESCC Low Expression 35865464 upregulation of circLONP2
    CRC tissue Low Expression 35837106 tumor tissue obteined from colorectal cancer patients
    HEK-293 Medium Expression 35861986
    PC3 Medium Expression 35710817
    A549 Low Expression 35668077
    WM3734 Tet3G-KDM5B Medium Expression 35650266
    BxPC3 Low Expression 35618735 C. BxPC-3 cells treated with recombinant human activin A (50 and 100 ng/ml) compared to untreated cells. e. pancreatic stellate cells (PSCs) proteins harvested from mono-cultured and direct co-cultured (D) tumor cells (BxPC-3-RFP) treated with IgG (1 μg/ml) for 48 h.
    RBE Low Expression 35517415
    9810 Low Expression 35517415
    SW480 Medium Expression 35414768
    SW620 Medium Expression 35414768
    HCT116 Low Expression 35372504
    HCT8 Low Expression 35372504
    HCMVECs Medium Expression 35571674
    NUGC4 Low Expression 35328684 rhHMGB 1（200 ng/mL，48 h）
    MKN7 Medium Expression 35328684 rhHMGB 1（200 ng/mL，48 h）
    U87MG Medium Expression 35317283
    A-172 Medium Expression 35317283
    U118MG Medium Expression 35317283
    P3 Low Expression 35317283
    U251 Low Expression 35317283
    LN229 Medium Expression 35317283
    LY1 Medium Expression 34743203
    LY3 Medium Expression 34743203
    A549 High Expression 35164653
    H1299 Medium Expression 35164653
    PANC1 Medium Expression 34998236
    AsPC1 No Expression 34998236
    BFTC909 Medium Expression 35205628
    AsPC1 Medium Expression 35127474
    BXPC3 High Expression 35127474
    HCC38 Low Expression 35027548 zeb1 protein enrichment in sorted αvβ3+ CSCs from HCC38 cells
    LM2-4 Medium Expression 35027548
    BT-549 Medium Expression 35027548 after treatment with rhTGFBI for 24 h with the indicated doses
    HCT116 Low Expression 35008299
    PANC1 Medium Expression 34513308
    BXPC3 Low Expression 34513308
    PC cells Low Expression 34513308 with conditioned medium from adipocytes
    UM-UC3 Medium Expression 34811353
    T24 Medium Expression 34811353
    A549 Low Expression 34884446 Cells were exposed to PM10 (10 µg/cm2) and TGF-β (5 ng/mL) for 48 h.
    JHH5 Low Expression 34772924
    HepG2 Low Expression 34772924
    Hep3B Medium Expression 34772924
    SNU449 High Expression 34772924
    MCF10A Low Expression 34674320
    U118MG Medium Expression 34595169
    HCT116 Medium Expression 34243012
    HaCaT No Expression 34466140
    NIH3T3 Low Expression 34466140
    PANC1 Low Expression 34522453 gemcitabine-resistant (GR) PC cell lines, PANC-1/GR and SW1990/GR.
    SW1990 Low Expression 34522453 gemcitabine-resistant (GR) PC cell lines, PANC-1/GR and SW1990/GR.
    HCT116 Low Expression 34502497
    T11 organoids (mice) Medium Expression 34253738
    T12 organoids (mice) Medium Expression 34253738
    MCF-7 Medium Expression 34008927
    HBL-100 High Expression 34141711
    MCF-7 No Expression 34141711
    T-47D No Expression 34141711
    BT-474 No Expression 34141711
    SK-BR-3 No Expression 34141711
    MDA-MB-453 No Expression 34141711
    MDA-MB-231 Low Expression 34141711
    HCC-1937 No Expression 34141711
    BT-549 No Expression 34141711
    HS-578-T Low Expression 34141711
    MDA-MB-468 No Expression 34141711
    4Z-B-1 No Expression 34141711
    Hep3B Low Expression 34093825
    Huh7 Medium Expression 34093825
    HCT116 Low Expression 34083586
    SW480 Medium Expression 34083586
    EMCs Medium Expression 34067060 treated with TGF β1 (1 ng/mL) for 24 h
    MDA-MB-231 Low Expression 33907253
    HS-578-T Low Expression 33907253
    GSC Low Expression 33894774 GSCs grown on NFSs with different stiffnesses.
    H1975 Medium Expression 33883697
    A549 Medium Expression 33883697 2 ng/ml TGF-β1 for 24 h
    NCI-H1975 No Expression 32678313 osimertinib-resistant NCI-H1975/OSIR cells.
    DU145 Low Expression 33542227
    C4-2 Medium Expression 33542227
    Cal62 Low Expression 33479369
    HTH83 Low Expression 33479369
    AS-B634 Medium Expression 33413566
    MDA-MB-231 Low Expression 33413566
    MDA-MB-231 Low Expression 33390847
    HCC-1937 Medium Expression 33390847
    SK-MEL-30 Low Expression 33291663 hGH (50 ng/ml、250 ng/ml) treatment for 48 h
    B16-F10 Medium Expression 33291663 bGH (500 ng/ml) treatment for 72 h
    CAOV3 No Expression 33188287
    OVCA 433 No Expression 33188287
    SK-OV-3 High Expression 33188287
    HEY High Expression 33188287
    HCT116 Medium Expression 33178684
    DLD-1 Low Expression 33178684
    SW1116 Medium Expression 33178684
    HT-29 Low Expression 33178684
    Huh7 Low Expression 32963356
    HGC-27 Low Expression 32963356
    PANC1 No Expression 33086527
    A549 Low Expression 32817494 A549 cells were treated with 5 ng/mL TGFβ1 to induce EMT for 3d 5d 7d 9d
    HGC-27 High Expression 32963944
    HCC827 Low Expression 32319651 gefitinib-resistant cell lines
    YD-10B Low Expression 32728068
    A549 Medium Expression 32546976
    H1299 Medium Expression 32546976
    786O Low Expression 32536815
    769P Low Expression 32536815
    MCF-7M Medium Expression 32396572
    JEG-3 No Expression 32319581 100 µM forskolin for 48 h at 37˚C.
    MCF-7 Low Expression 32227618
    MDA-MB-231 Medium Expression 32227618
    TPC1 Medium Expression 32157211
    Cal62 Medium Expression 32157211
    PCC13 Medium Expression 31846430
    K1 Medium Expression 31846430
    CAL27 No Expression 31924739
    FaDu No Expression 31924739
    Detroit-562 No Expression 31924739
    SCC25 No Expression 31924739
    SCC9 No Expression 31924739
    SCC15 No Expression 31924739
    DU145 High Expression 31685947
    22RV1 No Expression 31685947
    SiHa Medium Expression 31974393 acidosis-adapted cancer cells (cells were maintained in DMEM supplemented with 25 mmol/L of both PIPES and HEPES to adjust pH to 6.5.)
    FaDu Low Expression 31974393 acidosis-adapted cancer cells (cells were maintained in DMEM supplemented with 25 mmol/L of both PIPES and HEPES to adjust pH to 6.5.)
    HCT116 Low Expression 31974393 acidosis-adapted cancer cells (cells were maintained in DMEM supplemented with 25 mmol/L of both PIPES and HEPES to adjust pH to 6.5.)
    MG63 Medium Expression 31746422
    U2OS Medium Expression 31746422
    PHH Low Expression 31432507
    MIAPaCa2 Low Expression 31842285
    PANC1 Low Expression 31842285
    HCC827 No Expression 31811131
    SUNE1 Medium Expression 31433128
    CNE2 Low Expression 31433128
    BT-549 Medium Expression 31308076
    HBL-100 Low Expression 31462314
    MCF10A No Expression 31462314
    BT-474 No Expression 31462314
    MCF-7 No Expression 31462314
    MDA-MB-416 No Expression 31462314
    SK-BR-3 No Expression 31462314
    T-47D No Expression 31462314
    MDA-MB-453 No Expression 31462314
    ZR-75-1 No Expression 31462314
    ZR-75-30 No Expression 31462314
    HCC-1937 No Expression 31462314
    MDA-MB-468 No Expression 31462314
    BT-549 Medium Expression 31462314
    HS-578-T Medium Expression 31462314
    MDA-MB-231 Medium Expression 31462314
    MDA-MB-436 Low Expression 31462314
    Bcap-37 Low Expression 31462314
    SCC9 Medium Expression 31440468
    SAS Medium Expression 31440468
    PANC1 Medium Expression 31405071
    MIAPaCa2 Medium Expression 31405071
    LNCaP cells Low Expression 31379747 treated cells with 10 nM insulin for a further 48 h
    T24 Medium Expression  31268162
    human PAECs Medium Expression 31440380 exposed to TNFα (10 ng/ml) for 48 hours
    HCT116 Low Expression 31370857
    SW480 Medium Expression 31370857
    MDA-MB-231 Medium Expression 31181813
    SK-BR-3 Medium Expression 31181813
    U87 Low Expression 26553592
    U251 Medium Expression 26553592
    BCPAP Medium Expression 30814512
    TPC1 Medium Expression 30814512
    IGROV-1 Low Expression 30259564 incubation with CXCL12 (100 ng/mL) for 24 h
    HO-8910 Medium Expression 30259564 incubation with CXCL12 (100 ng/mL) for 24 h
    Huh7 Low Expression 30541499
    HME Low Expression 30220054
    MKN74 Low Expression 30473688
    NCI-N87 Low Expression 30473688
    PANC1 Low Expression 30156359 Panc‐1‐circulating tumor cells (CTC) cells, derived from Panc‐1‐parent (Panc‐1‐P) cells
    preSC Low Expression 30181244
    DMS53 Low Expression 30181244
    SW1990 Medium Expression 30066876
    PANC1 Medium Expression 29845225
    CFPAC-1 High Expression 29845225
    CFPAC-1 Medium Expression 29930735
    BXPC3 Medium Expression 29930735
    BCG823 Medium Expression 29367428
    MGC-803 Low Expression 29367428
    hTCEpi Low Expression 29747422
    501 mel No Expression 29369499
    SK-MEL-3 No Expression 29369499
    Hcclm3 High Expression 29286072
    HepG2 Medium Expression 29286072
    hTCEpi No Expression 29401709
    Tumor Medium Expression 29180466
    H1500 No Expression 29263324
    MCF-7 No Expression 29263324
    T47D No Expression 29263324
    ZR-75-1 No Expression 29263324
    DLD-1 No Expression 29263324
    HCT15 No Expression 29263324
    MDA-231 High Expression 29263324
    MDA-157 Medium Expression 29263324
    SUM149 No Expression 29263324
    HCT116 Low Expression 29263324
    SW480 Medium Expression 29263324
    A549 No Expression 29263324
    HESC Medium Expression 29116025
    NMuMG/E9 Low Expression 29079737 cultured in presence of TGFβ for 4 days
    HT-29 Medium Expression 28346422
    SW480 Medium Expression 28346422
    SW1116 Medium Expression 28346422
    MDA-MB-468 Medium Expression 28259959
    MDA-MB-231 Medium Expression 28259959
    HEK-293T Low Expression 27907888 treated 99021 (GSK3β inhibitor) (2 μM) for 72 h
    PANC1 High Expression 27878247
    BxPC3 Medium Expression 27878247
    NMuMG Low Expression 27617643 2 ng/ml TGF-β for 24 h or 48 h
    HCC827 No Expression 27456471
    H441 No Expression 27456471
    393P No Expression 27456471
    HCC827 No Expression 27456471
    H3255 No Expression 27456471
    H1299 Low Expression 27456471
    MDA-MB-231 Medium Expression 26387542
    Hela Medium Expression 26748910
    H1650 Medium Expression 26748910
    H1975 Low Expression 26748910
    H2030 Medium Expression 26748910
    PEO1 No Expression 26887977
    OVCAR5 Low Expression 26887977
    OVCA420 No Expression 26887977
    OVCA429 No Expression 26887977
    MDA-MB-231 Medium Expression 25849745
    SUM159 Low Expression 25849745
    PC3 Low Expression 25569101
    INS-1 Low Expression 25391656
    DLD-1 No Expression 25042806
    U2OS High Expression 25042806
    SJSA High Expression 25042806
    MIAPaCa2 Medium Expression 25042806
    PANC1 Medium Expression 25042806
    HCC827 No Expression 24762440
    H1299 Low Expression 24762440
    DU145 Low Expression 24659820 docetaxel-resistant cells
    PC3 No Expression 24659820 docetaxel-resistant cells
    UC6 Low Expression 23239884
    UC3 High Expression 23239884
    HAP1 Medium Expression
    Hela Medium Expression
    MDA-MB-231 High Expression
    SK-OV-3 Low Expression
    MCF-7 No Expression
    A549 No Expression
    human ovary lysates Medium Expression
    HEK-293 Medium Expression
    PLC/PRF/5 Medium Expression 36191855
    Huh7 Low Expression 36191855
    MHCC97L Low Expression 36202326
    PLC/PRF/5 Low Expression 36202326
    Hep3B Low Expression 36202326
    TS14-15 Medium Expression 37558923
    TS13-64 Medium Expression 37558923
    HCT116 Medium Expression 37810854
    HT-29 Medium Expression 37810854
    LN229 Medium Expression 37863960
    U251 Medium Expression 37863960
    EH-GB1 Medium Expression 34913073
    HPDE6C7 Low Expression 35186455
    AsPC1 Low Expression 35186455
    BXPC3 Medium Expression 35186455
    CFPAC-1 Medium Expression 35186455
    PANC1 Medium Expression 35186455
    HDMEC Medium Expression 35570385
    HONE1 Low Expression 35864158
    SUNE1 Low Expression 35864158
    U87MG Medium Expression 35111679
    U251 Medium Expression 35111679
    Hep-2 Medium Expression 34257531
    TU-177 Medium Expression 34257531
    FaDu Medium Expression 34853532
    Hep3B Medium Expression 32616906
    HepG2 Medium Expression 32616906
    T24 Medium Expression 33902589 10 ng/ml TGFβ1 (The processing time is not indicated)
    RT4 Low Expression 33902589 10 ng/ml TGFβ1 (The processing time is not indicated)
    HCCLM3 Medium Expression 33854606
    Huh7 Medium Expression 33854606
    HN3erPCC Medium Expression 33741422
    HN4erPCC Medium Expression 33741422
    A549 Medium Expression 33808696
    Pc9 Medium Expression 33808696
    Huh7 Medium Expression 33407720
    HPMC Medium Expression 32488650 IL-17A 100ng/ml（The processing time is not indicated）
    U251-MG Medium Expression 34422645
    U87MG Medium Expression 34422645
    AGS Low Expression 34405002
    MGC-803 Medium Expression 34405002
    HGC-27 Low Expression 34405002
    Hela Medium Expression 34493213
    C33A Medium Expression 34493213
    C33A xenograft Medium Expression 34493213
    SW620 Medium Expression 31755734
    SW480 Medium Expression 31755734
    A549 Medium Expression 33191397
    Pc9 Medium Expression 33191397
    7901 High Expression 32596004
    AGS Low Expression 32596004
    MCF-7 Medium Expression 33174044
    NB SK-N-SH Medium Expression 32793474
    SW780 Medium Expression 32862492
    T24 High Expression 32862492
    BXPC3 Low Expression 32945350 BxPC-3 cells were treated with TGF-β (5 ng/ml) for 28 days and were passaged every 3 days.
    PANC1 Low Expression 32945350
    AGS-EBC Medium Expression 33052232
    BRL-3A Low Expression 33058500 treated with Fructose (5 mM) for 1h 2h 4h
    Rat livers-nuclear、cytoplasm Low Expression 33058500 Each rat was given the drinking water or 100 mL drinking water containing 10% fructose (wt/vol) for 6 weeks。4 mg/kg pioglitazone
    BRL-3A Low Expression 33058500 5 mM Fructose and 10 μM pioglitazone
    AMC HN3 No Expression 32896720
    AMC HN4 No Expression 32896720
    AMC HN6 Medium Expression 32896720
    AMC HN7 No Expression 32896720
    AMC HN9 High Expression 32896720
    AMC HN10 Medium Expression 32896720
    AGS Low Expression 32303680
    SMMC7721 Medium Expression 32269215
    HCCLM3 Medium Expression 32269215
    U87MG Medium Expression 32368392
    U251 Medium Expression 32368392
    3T3 Medium Expression 31553422
    HT-29 Medium Expression 31164794
    HCT116 Medium Expression 31164794
    T24 Medium Expression 31115480
    UM-UC3 Medium Expression 31115480
    Caco-2 Low Expression 31281520
    DiFi Low Expression 31281520
    DLD-1 Medium Expression 31281520
    LoVo High Expression 31281520
    SK28 Medium Expression 31427543
    A375 Medium Expression 31427543
    LN18 Medium Expression 31429770
    U251 Medium Expression 31429770
    FARAGE Medium Expression 31570691
    U2932 Medium Expression 31570691
    IMR-32 High Expression
    Jurkat Medium Expression
    HT-1080 Low Expression
    A-172 Low Expression
    HCC1806 No Expression
    BA/F3 Low Expression
    YAC-1 Medium Expression
    HCC827 Low Expression 37239162 Osimertinib-resistant HCC827
    PC9 Medium Expression 37239162 Gefitinib-resistant PC9 (PC9/GR)
    T24 Low Expression 37086261
    UM-UC3 Medium Expression 37086261
    MCF10A Low Expression 36261627
    MCF-7 Low Expression 36302751 incubated with indicated exosomes
    MDA-MB-231 Medium Expression 36302751 incubated with indicated exosomes
    HCT116 Medium Expression 35978266
    SW480 Low Expression 35978266
    OB/OB mice Colorectal cancer tumor tissue Low Expression 35978266
    MCF-7 No Expression 35858297
    A549 Medium Expression 35790743
    H1299 Medium Expression 35790743
    Huh7 No Expression 35589690 cells pretreated with the CAF-CM (Cancer-associated fibroblasts - conditioned medium)
    Hep3B Low Expression 35589690 cells pretreated with the CAF-CM (Cancer-associated fibroblasts - conditioned medium)
    MGC-803 Medium Expression 35295342
    HGC-27 Medium Expression 35295342
    MDA-MB-231 Medium Expression 33664871
    MCF-7 Medium Expression 33664871
    HEK-293 No Expression
    HAP1 Low Expression
    MCF-7 Low Expression 34149926
    TAMR Medium Expression 34149926
    MKN28 Medium Expression 32020707
    MKN45 Medium Expression 32020707
    SK-BR-3 Medium Expression 32039833
    MDA-MB-435 Medium Expression 32039833
    MCF-7 Medium Expression 32039833
    HCC1937 Medium Expression 32039833
    HNSCC tissues Low Expression 33363013
    TPC1 High Expression 31257462
    HTH83 High Expression 31257462
    GES-1 Low Expression 31793989
    MGC-803 Medium Expression 31793989 Paclitaxel resistance
    AGS Medium Expression 31793989 Paclitaxel resistance
    PANC1 Medium Expression 30487699
    MIAPaCa2 Medium Expression 30487699
    Saos2 Medium Expression 28694763
    Click to view more sample data

    *For predicted expression levels of this protein in various human-derived cells and tissues, please refer to: http://www.proteinatlas.org

    Exprimental Methods

    WB
    Experimental Protocol:
     
    Sample preparation
    1. Tissue: Lyse the tissue sample by adding an appropriate volume of ice-cold RIPA/Nuclear Lysis Buffer (containing Protease Inhibitor Cocktail)，and homogenize the tissue at a low temperature or lyse it by sonication on ice, then incubate on ice for 30 minutes.
    2. Adherent cell: Aspirate the culture medium and transfer the cells into an EP tube. Wash the cells with ice-cold PBS twice. Add an appropriate volume of RIPA/Nuclear Lysis Buffer (containing Protease Inhibitor Cocktail), sonicate to lyse the cells, and incubate on ice for 30 minutes.
    3. Suspension cell: Transfer the culture medium to a pre-cooled centrifuge tube. Centrifuge and aspirate the supernatant. Wash the cells with ice-cold PBS twice.Add an appropriate volume of RIPA/Nuclear Lysis Buffer (containing Protease Inhibitor Cocktail), sonicate to lyse the cells, and incubate on ice for 30 minutes.
    4. Place the lysate into a pre-cooled microcentrifuge tube. Centrifuge at 4°C for 15 min. Collect the supernatant;
    5. Remove a small volume of lysate to determine the protein concentration;
    6. Combine the lysate with protein loading buffer. Boil 20 µL sample under 95-100°C for 5 min. Centrifuge for 5 min after cool down on ice.
     
    Electrophoretic separation
    1. According to the concentration of extracted protein, load appropriate amount of protein sample and marker onto SDS-PAGE gels for electrophoresis. Recommended separating gel (lower gel) concentration: 5%. Reference Table for Selecting SDS-PAGE Separation Gel Concentrations
    2. Power up 80V for 30 minutes. Then the power supply is adjusted (110 V~150 V), the Marker is observed, and the electrophoresis can be stopped when the indicator band of the predyed protein Marker where the protein is located is properly separated. (Note that the current should not be too large when electrophoresis, too large current (more than 150 mA) will cause the temperature to rise, affecting the result of running glue. If high currents cannot be avoided, an ice bath can be used to cool the bath.)
     
    Transfer membrane
    1. Take out the converter, soak the clip and consumables in the pre-cooled converter;
    2. Activate PVDF membrane with methanol for 1 min and rinse with transfer buffer;
    3. Install it in the order of "black edge of clip - sponge - filter paper - filter paper - glue -PVDF membrane - filter paper - filter paper - sponge - white edge of clip";
    4. The protein was electrotransferred to PVDF membrane. ( 0.45 µm PVDF membrane is recommended ) Reference Table for Selecting PVDF Membrane Pore Size Specifications
    Recommended conditions for wet transfer: 200 mA, 120 min.
    ( Note that the transfer conditions can be adjusted according to the protein size. For high-molecular-weight proteins, a higher current and longer transfer time are recommended. However, ensure that the transfer tank remains at a low temperature to prevent gel melting.)
     
    Block
    1. After electrotransfer, wash the film with TBST at room temperature for 5 minutes;
    2. Incubate the film in the blocking solution for 1 hour at room temperature;
    3. Wash the film with TBST for 3 times, 5 minutes each time.
     
    Antibody incubation
    1. Use 5% skim milk powder to prepare the primary antibody working liquid (recommended dilution ratio for primary antibody 1:1000), gently shake and incubate with the film at 4°C overnight;
    2. Wash the film with TBST 3 times, 5 minutes each time;
    3. Add the secondary antibody to the blocking solution and incubate with the film gently at room temperature for 1 hour;
    4. After incubation, wash the film with TBST 3 times for 5 minutes each time.
     
    Antibody staining
    521. Add the prepared ECL luminescent substrate (or select other color developing substrate according to the second antibody) and mix evenly;
    2. Incubate with the film for 1 minute, remove excess substrate (keep the film moist), wrap with plastic film, and expose in the imaging system.

    Datasheet & SDS

    Biological Description

    Specificity

    Anti-ZEB1 Rabbit Antibody [M1M18] detects endogenous levels of total TCF8/ZEB1 protein.
    Subcellular Location
    Nucleus
    Uniprot ID
    P37275
    Clone
    M1M18
    Background

    The ZEB family comprises zinc-finger and homeobox domain-containing transcription factors, with two members identified in mammals: ZEB1 (also known as δ-EF1, TCF8, AREB6) and ZEB2 (SIP1). Both ZEB1 and ZEB2 feature two distinct zinc-finger domains along with a homeodomain. While primarily acting as transcriptional repressors, ZEB proteins can also activate transcription, contingent upon the DNA context and cell type. Among their targets for repression is E-cadherin. The suppression of E-cadherin represents a key aspect of epithelial-mesenchymal transition (EMT), crucial for normal embryonic development and exploited by malignant epithelial tumors to metastasize from their tissue of origin. ZEB1 mutations are linked to posterior corneal dystrophy, whereas ZEB2 mutations have been associated with Hirschsprung (HSCR) disease.
    References
    • https://pubmed.ncbi.nlm.nih.gov/17486063/

    Tech Support

    Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

    Handling Instructions

    Tel: +1-832-582-8158 Ext:3
    If you have any other enquiries, please leave a message.

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