Home 一抗 Anti-ZEB1 Rabbit Antibody [M1M18] 仅限科研使用

Anti-ZEB1 Rabbit Antibody [M1M18]

目录号: F0266

    为确保有效检测，需先提取细胞核蛋白后再进行实验。如需获取不同样本的表达情况请点击“样品处理数据示例表”。
    抗体应用： 反应性：
    • Lane 1: Jurkat
      Lane 2: HT1080
      Lane 3: A172
      Lane 4: COS (mock transfected)
      Lane 5: COS (transfected with hZEB1)
    规格 价格 库存 购买数量
    20ul 790 现货
    50uL 1240 现货
    100ul 1990 现货
    100uL*2 3790 现货

    400-668-6834

    info@selleck.cn

    操作要点

    为确保有效检测，需先提取细胞核蛋白后再进行实验。

    WB
    建议 SDS-PAGE 分离胶浓度：5%。

    使用信息

    稀释比例
    1:1000
    1:200
    抗体应用
    WB, IP
    反应性
    Human
    抗体类型
    Rabbit
    储存液配方
    PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN₃
    储存条件(自收到货起)
    –20°C (avoid freeze-thaw cycles), 2 years
    预测分子量
    200 kDa
    阳性对照 COS (transfected with hZEB1); Jurkat; HT1080; A172
    阴性对照 COS

    样品处理数据示例

    细胞系 表达情况 参考文献 样本处理
    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
    点击查看更多样品数据

    *该蛋白在不同的人源细胞、组织内的表达量预测请参考：http://www.proteinatlas.org

    实验方法

    WB
    实验步骤：
     
    样品制备
    1. 组织样品：破碎组织，加入适量冰冷的 RIPA/Nuclear Lysis Buffer （含蛋白酶抑制剂Cocktail），低温匀浆或置于冰上超声裂解样品，静置30 min。
    2. 贴壁细胞样品：吸去培养基, 将细胞转移至EP管中。用冰冷的 PBS 清洗细胞 2 遍。加入适量冰冷的 RIPA/Nuclear Lysis Buffer （含蛋白酶抑制剂Cocktail），超声破碎，冰上静置裂解 30 min。冰上静置裂解 5 min。
    3. 悬浮细胞样品：将培养基转移至离心管中离心，弃上清，用冰冷的 PBS 清洗细胞 2 遍。加入适量冰冷的 RIPA/Nuclear Lysis Buffer （含蛋白酶抑制剂Cocktail），冰上静置裂解 5 min。
    4. 将所得匀浆液/裂解液置于离心机中 4°C 离心 15 min，收集上清液；
    5. 取少量裂解液测定蛋白质浓度；
    6. 加入蛋白上样缓冲液，将 20 µL样品在 95~100°C加热 5 min，冰上静置冷却后离心 5 min。
     
    电泳分离
    1. 根据所提蛋白的浓度，将适量蛋白样品和 Marker 上样至 SDS-PAGE 凝胶。建议分离胶（即下层胶）浓度：5 %。 SDS-PAGE 分离胶浓度选择参照表
    2. 电源调 80 V, 30 min。然后电源调 110 V~150 V，观察 Marker，待蛋白所在的预染蛋白 Marker 指示带得到合适的分离后，即可停止电泳。（注意电泳时电流切勿过大，如超过 150 mA 会导致温度上升，容易影响跑胶结果。如无法避免采用大电流，可对电泳槽使用冰浴降温。）
     
    转膜
    1.拿出电转槽，把夹子和耗材浸泡在预冷电转液中；
    2.用甲醇活化 PVDF 膜 1 min，用转移缓冲液冲洗；
    3.按照“夹子黑边-海绵-滤纸-滤纸-胶-PVDF膜-滤纸-滤纸-海绵-夹子白边”的顺序安装好；
    4.将蛋白电转移至 PVDF膜上。（推荐采用 0.45 µm PVDF 膜） PVDF 膜孔径规格选择参照表
    湿转法参考条件：200 mA, 120 min。
    （注意电转条件可根据蛋白大小适当调节，分子量大的蛋白适宜采用大电流，并延长转膜时间，但需要确保电转槽始终处于低温的环境中，避免凝胶融化。）
     
    封闭
    1. 电转移后，室温下用 TBST 洗膜 5 min；
    2. 在封闭液中将膜孵育 1 h，室温；
    3. 用 TBST 洗膜 3 次，每次 5 min。
     
    抗体孵育
    1. 用 一抗稀释液配制一抗工作液（建议一抗稀释比 1:1000），4°C 条件下与膜轻柔摇晃孵育过夜；
    2. 用 TBST 洗膜 3 次，每次 5 min；
    3. 在封闭液中加入二抗，室温条件下与膜轻柔摇晃孵育 1 h；
    4. 孵育结束后，用 TBST 洗膜 3 次，每次 5 min。
     
    显色
    1. 加入配制好的 ECL 发光底物（或根据二抗选择其他显色基质）混合均匀；
    2. 与膜孵育1 min，去除多余底物（需保持膜湿润），置于显影仪中进行曝光。
     

    Datasheet & SDS

    生物描述

    特异性

    Anti-ZEB1 Rabbit Antibody [M1M18] 可检测 TCF8/ZEB1 总蛋白的内源水平。
    蛋白定位
    细胞核
    Uniprot ID
    P37275
    克隆号
    M1M18
    背景

    ZEB 家族包含含有锌指和同源框结构域的转录因子，在哺乳动物中发现了两个成员：ZEB1（也称为 δ-EF1、TCF8、AREB6）和 ZEB2 (SIP1)。 ZEB1 和 ZEB2 均具有两个不同的锌指结构域和一个同源结构域。 虽然 ZEB 蛋白主要充当转录抑制因子，但它也可以激活转录，具体取决于 DNA 背景和细胞类型。 他们的镇压目标之一是E-钙粘蛋白。 E-钙粘蛋白的抑制代表了上皮-间质转化 (EMT) 的一个关键方面，对于正常胚胎发育至关重要，并被恶性上皮肿瘤利用从其起源组织转移。 ZEB1 突变与后角膜营养不良有关，而 ZEB2 突变与先天性巨结肠 (HSCR) 病有关。
    参考文献
    • https://pubmed.ncbi.nlm.nih.gov/17486063/

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