蛋白免疫印迹一站式解决方案
One-Stop Solution for Western Blot

蛋白质印迹法 (WB) 是一种广泛使用的基于抗体的技术，用于检测细胞或组织提取物中的蛋白质表达水平。这项技术借助抗体与目的蛋白的结合作用，测量生物样品中的蛋白质水平。

  • 1. 样本制备与定量
  • 2. 电泳与转印
  • 3. 封闭与抗体孵育
  • 4. 发光和成像

样本制备是Western Blot实验的基石，其质量直接决定后续步骤的成败。此阶段首先需要从细胞或组织中提取总蛋白。常用的裂解液含有去垢剂、盐类和蛋白酶抑制剂，旨在高效溶解蛋白质，同时防止蛋白降解。提取后，需要通过BCA法、Bradford法等对蛋白质样本进行精确定量，以确保后续上样量的一致性和结果的可比性。随后，将定量后的蛋白质与上样缓冲液混合，并在高温下变性，使蛋白质线性化并带上负电荷，为下一步的电泳分离做好准备。

试剂

实验流程产品名称备注货号
样本制备Bovine Serum Albumin (BSA)牛血清白蛋白，粉末RM02802
样本制备PBS Buffer Powder {PBS缓冲液粉剂(1*)}配置用的常规PBS，不是细胞培养级RM00012
样本制备Cell lysis buffer for IP (without inhibitors)支持WB、IP、co-IP实验，细胞裂解液RM00022
样本制备SDS-PAGE Sample Loading Buffer (5*)样品上样loadingRM00001
样本制备蛋白酶抑制剂Cocktail (不含EDTA，100X DMSO储液)蛋白酶抑制剂，抑制样品蛋白降解RM02916
样本制备Phosphatase Inhibitor Cocktail (2 Tubes, 100X)磷酸酶抑制剂，维持样品磷酸化状态，磷酸化检测必备RM02997
样本制备PMSF {苯甲基磺酰氟} (100mM)丝氨酸蛋白酶、巯基蛋白酶抑制剂，一般搭配裂解液使用RM70000
样本制备RIPA Lysis Buffer (Strong)强RIPA裂解液（裂解、变性程度不同，无特殊需求就选强）RM02998
样本制备RIPA Lysis Buffer (Medium)中RIPA裂解液（裂解、变性程度不同，无特殊需求就选强）RM02999
样本制备RIPA Lysis Buffer (Weak)弱RIPA裂解液（裂解、变性程度不同，无特殊需求就选强）RM03000
样本制备Recombinant Serratia marcescens Nuclease降解样品核酸杂质，提升WB样本质量RPT0008LQ
蛋白定量BCA Protein Assay Kit {BCA 蛋白定量试剂盒}测定蛋白浓度，确保各样本上样量一致BRK0011

仪器设备

高效组织细胞样品处理系统

  • 高效：三维振动，6000rpm最高转速，20s裂解完成95%的样品
  • 便捷：LED触摸屏，不需其他附加设备
  • 安全：紧急停止按钮

高效低温组织研磨仪

  • 配备不同适配器，最高可同时处理192样品
  • 支持最低-50°处理样本
  • 搭配不同适配器，满足各种需求

电泳与转印是WB实验的核心分离与转移步骤。电泳通常在聚丙烯酰胺凝胶（SDS-PAGE）中进行。在电场作用下，变性的蛋白质根据其分子量大小在凝胶中迁移，分子量小的跑得快，分子量大的跑得慢，从而实现蛋白质的精细分离。电泳结束后，需要将凝胶中分离好的蛋白质条带转移到固相支持膜（常用PVDF膜或NC膜）上，以便进行后续的抗体检测。转印在电场中进行，目的是让蛋白质从凝胶中“印迹”到膜表面，并牢固地结合在上面。这个过程确保了蛋白质的位置分布被固定下来，为免疫检测创造了条件。

试剂

实验流程产品名称备注货号
电泳One-Step Color Fast Gel Preparation Kit (6%) {一步法彩色快速凝胶试剂盒(6%)}制胶试剂盒，预混液，快速一步制胶BRK0001
电泳One-Step Color Fast Gel Preparation Kit (8%) {一步法彩色快速凝胶试剂盒(8%)}制胶试剂盒，预混液，快速一步制胶BRK0002
电泳One-Step Color Fast Gel Preparation Kit (10%) {一步法彩色快速凝胶试剂盒(10%)}制胶试剂盒，预混液，快速一步制胶BRK0003
电泳One-Step Color Fast Gel Preparation Kit (12%) {一步法彩色快速凝胶试剂盒(12%)}制胶试剂盒，预混液，快速一步制胶BRK0004
电泳One-Step Color Fast Gel Preparation Kit (15%) {一步法彩色快速凝胶试剂盒(15%)}制胶试剂盒，预混液，快速一步制胶BRK0005
电泳Precast Tris-Gly (10%)10 well Ultra-Fast Protein Electrophoresis Kit {预制Tris-Gly (10%) 10孔超快蛋白电泳试剂盒}10孔， 梯度预制胶+配套电泳液，Tris-gly体系BRK1010T
电泳Precast Tris-Gly (12%)10 well Ultra-Fast Protein Electrophoresis Kit {预制Tris-Gly (12%) 10孔超快蛋白电泳试剂盒}10孔， 梯度预制胶+配套电泳液，Tris-gly体系BRK1210T
电泳Precast Tris-Gly (4-20%)10 well Gradient Gel Ultra-Fast Protein Electrophoresis Kit {预制Tris-Gly (4-20%) 10孔超快蛋白电泳试剂盒}10孔， 梯度预制胶+配套电泳液，Tris-gly体系BRK0021P
电泳Precast Bis-Tris (10%) 10 well Protein Electrophoresis Kit｛预制Bis-Tris（10%）10孔蛋白电泳试剂盒｝10孔，梯度预制胶+配套电泳液, Bis-Tris体系BRK1010B
电泳Precast Bis-Tris (12%) 10 well Protein Electrophoresis Kit｛预制Bis-Tris（12%）10孔蛋白电泳试剂盒｝10孔，梯度预制胶+配套电泳液, Bis-Tris体系BRK1210B
电泳Precast Bis-Tris (4-20%) 10 well Gradient Gel Protein Electrophoresis Kit｛预制Bis-Tris（4-20%）10孔蛋白电泳试剂盒｝10孔，梯度预制胶+配套电泳液, Bis-Tris体系BRK0031
电泳Precast Tris-Gly (4-20%)15 well Gradient Gel Ultra-Fast Protein Electrophoresis Kit {预制Tris-Gly (4-20%) 15孔超快蛋白电泳试剂盒}15孔，梯度预制胶+配套电泳液，Tris-gly体系BRK0020P
电泳Precast Bis-Tris (4-20%) 15 well Gradient Gel Protein Electrophoresis Kit｛预制Bis-Tris（4-20%）15孔蛋白电泳试剂盒｝15孔，梯度预制胶+配套电泳液，Bis-Tris体系BRK0030
电泳Stain-Free Precast Tris-Gly (4-20%)15 well Gradient Gel Ultra-Fast Protein Electrophoresis Kit {预制免染Tris-Gly ,4-20%,15孔超快蛋白电泳试剂盒}15孔, 免染版的Tris-Gly梯度预制胶BRK0020TV
电泳Stain-Free Precast Tris-Gly (4-20%)10 well Gradient Gel Ultra-Fast Protein Electrophoresis Kit {预制免染Tris-Gly,4-20%,10孔超快蛋白电泳试剂盒}10孔, 免染版的Tris-Gly梯度预制胶BRK0021TV
电泳Stain-Free Precast Bis-Tris (4-20%) 15 well Gradient Gel Ultra-Fast Protein Electrophoresis Kit {预制免染Bis-Tris,4-20%,15孔超快蛋白电泳试剂盒}15孔, 免染版的Bis-Tris梯度预制胶BRK0030BV
电泳Stain-Free Precast Bis-Tris (4-20%)10 well Gradient Gel Ultra-Fast Protein Electrophoresis Kit {预制免染Bis-Tris,4-20%,10孔超快蛋白电泳试剂盒}10孔, 免染版的Bis-Tris梯度预制胶BRK0031BV
电泳ColorMixed Protein Marker 250 (10-250 kDa) {蛋白marker} 预染彩色蛋白markerRM02949
电泳ColorMixed Protein Marker 180 (10-180 kDa) {蛋白marker}预染彩色蛋白marker（10-180 kDa，10个带）RM19001
电泳ColorMixed Protein Marker 180 (10-180 kDa) Plus {蛋白marker}预染彩色蛋白marker（10-180 kDa，11个带）RM19001P
电泳ColorMixed Protein Marker 310 (10-310 kDa) {蛋白marker} 预染彩色蛋白markerRM02943
电泳Tris-Glycine-SDS Running Buffer Powder {Tris-glycine-SDS 电泳缓冲液粉剂1*}1*蛋白变性（含SDS）电泳液RM00010
电泳Tris-Glycine-Native Electrophoresis Buffer Powder｛Tris-Gly-Native电泳缓冲液粉末｝1*蛋白非变性（不含SDS）电泳液BR00006
电泳Ultra-Fast Tris-Glycine SDS Electrophoresis Buffer Powder｛Tris-Gly SDS 超快电泳缓冲液粉末｝超快Tris-Gly电泳体系，电泳超快，可搭配上面快速凝胶试剂盒BR00007
电泳Tris-Mops-SDS Electrophoresis Buffer Powder｛Tris-Mops-SDS电泳缓冲液粉末 适用Bis-Tris胶｝Tris-Mops电泳体系，可搭配上面快速凝胶试剂盒BR00008
电泳Ice-Bath Free Ultra-Fast Transfer Buffer Powder {免冰浴超快转膜缓冲液粉末}免冰浴超快转膜BR00009
电泳Decolorization-free Ultra Fast Staining Buffer考马斯亮蓝快速染色液 (免脱色) 考马斯亮蓝快速染色液（免脱色）BR00017
电泳Ponceau S Stain Solution 1X {丽春红染色液,1X}丽春红染色液,1XBR00036
转印Western Blot Stripping buffer 抗体洗脱液（WB专用）抗体洗脱液（再生液、剥离液）BR00016
转印Polyvinylidene fluoride (0.22μm) {PVDF膜 (6.0×9.0cm/pcs)(30cm*300cm/1卷) (0.22μm)}国产聚偏二氟乙烯膜，适合转小分子蛋白（高灵敏度和高分辨率，需甲醇活化）BR00102
转印Polyvinylidene fluoride (0.45μm) {PVDF膜 (6.0×9.0cm/pcs)(30cm*300cm/1卷) (0.45μm)}国产聚偏二氟乙烯膜，适合转大小分子蛋白（高灵敏度和高分辨率，需甲醇活化）BR00103
转印Nitrocellulose membrane (0.22μm) {NC膜 (6.0×9.0cm/pcs)(30cm*300cm/1卷) (0.22μm)}国产硝酸纤维素膜，适合转小分子蛋白（预算有限，灵敏度不高，操作简单无需预处理）BR00106
转印Nitrocellulose membrane (0.45μm) {NC膜 (6.0×9.0cm/pcs)(30cm*300cm/1卷) (0.45μm)}国产硝酸纤维素膜，适合转大小分子蛋白（预算有限，灵敏度不高，操作简单无需预处理）BR00107

仪器设备

电泳槽

  • 可四块胶同时运行
  • 封边隔条使得玻璃板加厚，不易破碎优势

转印槽

  • 支持高电流快速转印或低电流过夜转印
  • 内置2个冷冻模块，可快速轮换吸收缓冲液热量
  • 可快速高质地将蛋白样品转移到硝酸纤维膜、PVDF膜等介质上，可同时放置1-4个转印夹

电源

  • 四组并联，可同时带多个电泳槽优势
  • 液晶屏同时显示电压、电流、功率、定时时间
  • 安全性能：过压、电弧、空载和荷载突变监测；过载/短路监测；漏电保护；开路报警，断电自动恢复，暂停/恢复功能

全自动蛋白印记系统

  • 高通量：最多可支持12个通道同时工作
  • 全自动：封闭、孵育、清洗和一抗二抗回收全程自动，程序结束自动清洗管路
  • 高灵活：各通道程序独立控制，4种不同处理模式，不同环节可设置不同温度，多种尺寸孵育盒可选

转印后的膜上还有许多非特异性蛋白结合位点，如果不进行封闭，后续加入的抗体会非特异性地结合到这些位点，导致高背景和假阳性。因此，首先需要用脱脂牛奶或BSA等封闭液进行处理，这些蛋白质会占据膜上的非特异性位点。随后进行抗体孵育：先加入能特异性识别目标蛋白的一抗，使其与膜上的目标蛋白结合；洗涤去除未结合的一抗后，再加入带有标记物（如HRP酶）的二抗，二抗能特异性地识别并结合一抗。通过这种“抗体-抗原-抗体”的级联放大作用，最终使目标蛋白信号能够被检测到。

WB验证一抗

抗体选择指南

在进行 Western blot 实验时，使用内参蛋白是为了确保各个样品的上样量一致，并验证不同样品中目标蛋白表达变化的真实性。 这在检测阴性样本时尤其重要，例如基因敲除细胞系或用 siRNA 处理以敲低基因表达的细胞。 良好的内参对照可让您确信阴性样本是真实的阴性。 从而保证您的研究成果达到同行评审期刊发表的标准，并使您的工作能够被其他研究人员成功重现。

适用于多物种超高稀释比1:256W爆款内参β-Actin和GAPDH
β-Actin 重组兔单抗文献引用 : 2584

AC026

β-Actin Rabbit mAb (High Dilution)

样本：HeLa cells
  • 应用：WB,IF/ICC,IHC-P,ELISA
  • 物种：Human, Mouse, Rat, Chicken, Zebrafish, Pig, Cow
GAPDH 重组兔单抗文献引用 : 466

A19056

GAPDH Rabbit mAb(High Dilution)

样本：HeLa cells
  • 应用：WB,IHC-P,IF/ICC,ELISA
  • 物种：Human, Mouse, Rat
GAPDH 鼠单抗文献引用 : 300

AC033

GAPDH Mouse mAb (High Dilution)

样本：HeLa cells
  • 应用：WB,IHC-P,IF/ICC,ELISA
  • 物种：Human, Mouse, Rat
细胞核常见内参，应用物种全面超高稀释比1:128W爆款内参Histone H3、Lamin-B1和PCNA
Lamin-B1重组兔单抗

AC057

Lamin-B1 Rabbit mAb

人结肠组织
IHC稀释比1:16000
  • 应用：WB,IHC-P,IF/ICC,IP,ELISA
  • 物种：Human, Mouse, Rat
Histone H3重组兔单抗

AC070

Histone H3 Rabbit mAb

HeLa cells大鼠肺组织
IHC稀释比1:5500
  • 应用：WB,IP,IHC-P,ChIP,ELISA
  • 物种：Human, Mouse, Rat, Other (Wide Range Predicted)
PCNA鼠单抗

AC058

PCNA Mouse mAb

大鼠小肠组织
IF-P稀释比1:200
  • 应用：WB,IF/ICC,IF-P,IHC-P,ELISA
  • 物种：Human, Mouse, Rat
常见内参抗体清单
全细胞和细胞质
靶点货号名称应用物种文献数量
ACTBAC026β-Actin Rabbit mAb (High Dilution)WB,IF/ICC,IHC-P,ELISAHuman, Mouse, Rat, Chicken, Zebrafish, Pig, Cow2584
ACTBAC004β-Actin Mouse mAbWB,IHC-P,IF/ICC,ELISAHuman, Mouse, Rat443
ACTBAC038β-Actin Rabbit mAbWB,IHC-P,ELISAHuman, Mouse, Rat, Chicken, Zebrafish, Pig224
ACTBAC050β-Actin Rabbit mAbWB,IHC-P,IF/ICC,ELISAHuman, Mouse, Rat5
ACTBAC043HRP-conjugated β-Actin Mouse mAbWB,ELISAHuman, Mouse, Rat, Monkey5
ACTBAC028HRP-conjugated β-Actin Rabbit mAbWB,ELISAHuman, Mouse, Rat, Chicken, Zebrafish, Pig77
ACTBAC009Actin (plant specific) Mouse mAbWB,ELISAVarious Plants99
GAPDHAC033GAPDH Mouse mAb (High Dilution)WB,IHC-P,IF/ICC,ELISAHuman, Mouse, Rat300
GAPDHAC002GAPDH Mouse mAbWB,IHC-P,IF/ICC,ELISAHuman, Mouse, Rat837
GAPDHA19056GAPDH Rabbit mAb(High Dilution)WB,IHC-P,IF/ICC,ELISAHuman, Mouse, Rat466
GAPDHAC036GAPDH (Human Specific) Rabbit mAbWB,IF/ICC,ELISAHuman21
GAPDHAC054HRP-conjugated GAPDH Rabbit mAbWB,ELISAHuman, Mouse, Rat1
GAPDHAC035HRP-conjugated GAPDH Mouse mAbWB,ELISAHuman, Mouse, Rat37
TUBA4AAC012α-Tubulin Mouse mAbWB,IHC-P,IF/ICC,ELISAHuman, Mouse, Rat107
TUBA1BAC049α-Tubulin Rabbit mAbWB,IF/ICC,IHC-P,ELISAHuman, Mouse, Rat1
TUBBAC021β-Tubulin Mouse mAbWB,IHC-P,IF/ICC,IP,ELISAHuman, Mouse, Rat, Hamster, Monkey, Goat, Chlamydomonas Reinhardtii86
TUBBA12289β-Tubulin Rabbit mAbWB,IHC-P,IP,ELISAHuman, Mouse, Rat52
TUBBAC030HRP-conjugated β-Tubulin Mouse mAbWB,ELISAHuman, Mouse, Rat25
VCLA27905[KD Validated] Vinculin Rabbit mAbWB,IF-P,IHC-P,ELISAHuman, Mouse, Rat0
细胞膜
靶点货号名称应用物种文献数量
ATP1AA0643ATP1A Rabbit pAbWB,IHC-P,IP,ELISAHuman, Mouse4
CAV1A22417Caveolin-1 Rabbit mAbWB,IF/ICC,IHC-P,IP,ELISAHuman, Mouse, Rat1
CAV1A19006[KO Validated] Caveolin-1 Rabbit mAbWB,IHC-P,ELISAHuman, Mouse, Rat13
ATP1A1A11683Na+/K+-ATPase Rabbit mAbWB,IF/ICC,IF-P,IHC-P,ELISAHuman, Mouse, Rat14
细胞核
靶点货号名称应用物种文献数量
HDAC1A26492[KO Validated] HDAC1 Rabbit mAbWB,IP,IF/ICC,IF-P,IHC-P,ChIP,ELISAHuman, Mouse0
LMNAA19524[KO Validated] Lamin A/C Rabbit mAbWB,IHC-P,IF/ICC,IP,ELISAHuman, Mouse, Rat19
H3C1/H3-4AC070Histone H3 Rabbit mAbWB,IP,IHC-P,ChIP,ELISAHuman, Mouse, Rat, Other (Wide Range Predicted)0
LMNB1AC057Lamin-B1 Rabbit mAbWB,IHC-P,IF/ICC,IP,ELISAHuman, Mouse, Rat0
PCNAAC058PCNA Mouse mAbWB,IF/ICC,IF-P,IHC-P,ELISAHuman, Mouse, Rat0
YY1A19569YY1 Rabbit mAbWB,IF/ICC,ELISA,ChIPHuman, Mouse, Rat4
线粒体
靶点货号名称应用物种文献数量
VDAC1A19707VDAC1 Rabbit mAbWB,IHC-P,IF/ICC,ELISAHuman, Mouse, Rat39
SDHAA13852SDHA Rabbit mAbWB,IF/ICC,IF-P,ELISAHuman, Mouse, Rat1
COX4I1A28231[KD Validated] COX IV Rabbit mAbWB,IP,IF/ICC,IHC-P,ELISAHuman, Mouse, Rat0
HSPD1A0564HSP60/HSPD1 Rabbit mAbWB,IHC-P,IF/ICC,ELISAHuman, Mouse, Rat, Wheat11
血清
靶点货号名称应用物种文献数量
TFA19130Transferrin Rabbit mAbWB,IHC-P,ELISAHuman7

抗标签抗体是高度特异性的抗体，可用于定位与标签融合的蛋白。标签可帮助纯化蛋白质，还可用于研究尚无可用抗体的新型蛋白。 常见标签包括 DDDDK、GST、His、Myc 等。

IP小能手，探寻蛋白“好朋友”多应用，超高稀释比爆款标签，任您选择
磁珠DDDDK-Tag 鼠单抗文献引用 : 6

AE037

Magnetic Beads-conjugated Mouse anti DDDDK-Tag mAb

  • 应用：IP
  • 物种：Species independent
  • 样本：293T cells transfected with a SERPINB1 expression vector containing a single N-terminal DDDDK-Tag
DDDDK-Tag 重组兔单抗文献引用 : 50

AE092

DDDDK-Tag Rabbit mAb

  • 应用：WB,IF/ICC,IP,FC,ELISA,ChIP,ChIP-seq
  • 物种：Species independent
  • 样本：293T cells transfected with a SERPINB1 expression vector containing a single N-terminal DDDDK-Tag
DDDDK-Tag 鼠单抗文献引用 : 281

AE005

Mouse anti DDDDK-Tag mAb

  • 应用：WB,IF/ICC,IP,FC (intra),ELISA
  • 物种：Species independent
  • 样本：293T cells transfected with a SERPINB1 expression vector containing a single N-terminal DDDDK-Tag
常见标签抗体清单
常见标签抗体
靶点货号名称来源应用文献数量
DDDDKAE005Mouse anti DDDDK-Tag mAbMouseWB,IF/ICC,IP,FC (intra),ELISA281
DDDDKAE092DDDDK-Tag Rabbit mAbRabbitWB,IF/ICC,IP,FC,ELISA,ChIP,ChIP-seq50
GFPAE012Mouse anti GFP-Tag mAbMouseWB,IP,IF/ICC246
GFPAE078Rabbit anti GFP-Tag mAbRabbitWB,IF/ICC,IP,ELISA,ChIP14
GSTAE001Mouse anti GST-Tag mAbMouseWB,IP,ELISA104
GSTAE077Rabbit anti GST-Tag mAbRabbitWB,IP,ELISA6
HAAE008Mouse anti HA-Tag mAbMouseWB,IP,IF/ICC,ELISA226
HAAE105HA-Tag Rabbit mAbRabbitWB,IF/ICC,IP,ELISA13
HisAE003Mouse anti His-Tag mAbMouseWB,IF/ICC,IP,ELISA224
HisAE086Rabbit anti His-tag mAbRabbitWB,IF/ICC,IP,ELISA15
MBPAE016Mouse anti MBP-Tag mAbMouseWB,ELISA44
mCherryAE002Mouse anti mCherry-Tag mAbMouseWB,ELISA30
mCherryAE127Rabbit anti mCherry-Tag mAbRabbitWB,IP,ELISA1
MycAE010Mouse anti Myc-Tag mAbMouseWB,IP,IF/ICC,ELISA136
MycAE070Myc-Tag Rabbit mAbRabbitWB,IF/ICC,IP,ELISA76
Strep IIAE066Mouse anti Strep II-Tag mAbMouseWB,ELISA25
TAPAE021Mouse anti TAP-Tag mAbMouseWB,ELISA1
TRXAE023Mouse anti Trx-Tag mAbMouseWB,ELISA1
V5AE017Mouse anti V5-Tag mAbMouseWB,IP,IF/ICC,ELISA17
V5AE089Rabbit anti V5-Tag mAbRabbitWB,IF/ICC,ELISA,ChIP3
偶联标签抗体
靶点货号名称来源应用文献数量
DDDDKAE061Affinity Gel-conjugated Mouse anti DDDDK-Tag mAb {Anti-DDDDK Affinity Gel}MouseIP4
DDDDKAE121Affinity Gel-conjugated Rabbit anti DDDDK-Tag mAb {Anti-DDDDK Affinity Gel}RabbitIP3
DDDDKAE024HRP-conjugated Mouse anti DDDDK-Tag mAbMouseWB,ELISA16
DDDDKAE037Magnetic Beads-conjugated Mouse anti DDDDK-Tag mAbMouseIP6
GFPAE112Affinity Gel-conjugated Mouse anti GFP-Tag mAb {Anti-GFP Affinity Gel}MouseIP0
GFPAE119Affinity Gel-conjugated Rabbit anti GFP-Tag mAb {Anti-GFP Affinity Gel}RabbitIP1
GFPAE030HRP-conjugated Mouse anti GFP-Tag mAbMouseWB,ELISA1
GFPAE129HRP-conjugated Rabbit anti GFP-Tag mAbRabbitWB,ELISA0
GFPAE031HRP-conjugated Rabbit anti GFP-Tag mAbRabbitWB,ELISA1
GFPAE168Magnetic Beads-conjugated Mouse anti GFP-Tag mAbMouseIP0
GFPAE118Magnetic Beads-conjugated Rabbit anti GFP-Tag mAbRabbitIP1
GSTAE027HRP-conjugated Mouse anti GST-Tag mAbMouseWB4
GSTAE122Magnetic Beads-conjugated Rabbit anti GST-Tag mAbRabbitIP0
HAAE059Affinity Gel-conjugated Mouse anti HA-Tag mAb {Anti-HA Affinity Gel}MouseIP1
HAAE025HRP-conjugated Mouse anti HA-Tag mAbMouseWB,ELISA6
HAAE123HRP-conjugated Rabbit anti HA-Tag mAbRabbitWB0
HisAE028HRP-conjugated Mouse anti His-Tag mAbMouseWB,ELISA30
HisAE104HRP-conjugated Rabbit anti His-Tag mAbRabbitWB,ELISA2
MBPAE075HRP-conjugated Mouse anti MBP-TagMouseWB2
MycAE120Affinity Gel-conjugated Rabbit anti Myc-Tag mAb {Anti-Myc-Tag Affinity Gel}RabbitIP2
MycAE026HRP-conjugated Mouse anti Myc-Tag mAbMouseWB,ELISA5
MycAE102HRP-conjugated Rabbit anti Myc-Tag mAbRabbitWB,ELISA0
V5AE029Agarose Beads-conjugated Mouse anti V5-Tag mAbMouseIP0
对照IgG
靶点货号名称来源应用文献数量
Control IgGAC005Rabbit Control IgGRabbitIP,ChIP237
Control IgGAC011Mouse Control IgGMouseIP98
IgG isotype controlAC042Rabbit IgG isotype controlRabbitWB,IHC-P,IF/ICC,FC,ChIP,ELISA9

单域抗体(Single Domain Antibody,SdAb)，又称纳米抗体(Nanobody)或者VHH抗体(Variable Domain of Heavy Chain of Heavy-chain Antibody)，只含一条多肽链，是目前发现的能与抗原特异结合的最小抗体(约15KD)。基于纳米抗体技术，以及纳米抗体具有良好的溶解性，对热、蛋白酶和pH值极其稳定，ABclonal推出了GFP、mCherry和DDDDK纳米抗体Beads。

GFP纳米抗体Beads文献引用 : 14

AE079

Magnetic beads-conjugated anti-GFP VHH Single Domain antibody

  • 应用：IP,ChIP,CoIP,RIP
  • 物种：Species independent
  • 样本：293T cells transfected with a SERPINB1 expression vector containing a single C-terminal GFP-Tag
mCherry纳米抗体Beads文献引用 : 2

AE094

Magnetic beads-conjugated anti-mCherry VHH Single Domain antibody

  • 应用：IP,ChIP,CoIP,RIP
  • 物种：Species independent
  • 样本：293T cells transfected with a CT55 expression vector containing a single C-terminal mCherry-Tag
DDDDK纳米抗体Beads文献引用 : 1

AE126

Magnetic Beads-conjugated anti-DDDDK tag VHH Single Domain antibody

  • 应用：IP
  • 物种：
  • 样本：293T cells transfected with GSK3B-DDDDK-Tag
纳米抗体清单
靶点货号名称来源应用文献数量
DDDDKAE125Agarose beads-conjugated anti-DDDDK tag VHH Single Domain antibodyAlpacaIP0
DDDDKAE126Magnetic Beads-conjugated anti-DDDDK tag VHH Single Domain antibodyAlpacaIP1
mCherryAE073Agarose beads-conjugated anti-mCherry VHH Single Domain antibodyAlpacaIP,ChIP,CoIP1
mCherryAE094Magnetic beads-conjugated anti-mCherry VHH Single Domain antibodyAlpacaIP,ChIP,CoIP,RIP2
GFPAE074Agarose beads-conjugated anti-GFP VHH Single Domain antibodyAlpacaIP,ChIP,CoIP,RIP17
GFPAE079Magnetic beads-conjugated anti-GFP VHH Single Domain antibodyAlpacaIP,ChIP,CoIP,RIP14
MycAE106Agarose beads-conjugated anti-Myc VHH Single Domain antibodyAlpacaIP0
MycAE107Magnetic beads-conjugated anti-Myc VHH Single Domain antibodyAlpacaIP0
HAAE108Agarose beads-conjugated anti-HA VHH Single Domain antibodyAlpacaIP0
HAAE109Magnetic beads-conjugated anti-HA VHH Single Domain antibodyAlpacaIP0

“二抗”是第二抗体的简称，在Western blot中广泛使用。它的主要作用是识别并结合一抗（第一抗体），通常带有可被检测的标记（如酶、荧光素、生物素等），从而实现对目标抗原的间接检测。

HRP偶联羊抗兔二抗文献引用 : 1998

AS014

HRP-conjugated Goat anti-Rabbit IgG (H+L)

  • 应用：WB,DB,IHC-P,ELISA
  • 物种：Rabbit
  • Western blot analysis of lysates from HeLa cells,using GAPDH(AC001)antibody as the primary antibody at dilution of 1:80000.
    Secondary antibody:using HRP Goat Anti-Rabbit IgG (H+L)antibody (AS014)at 1:4000-1:10000 dilution.
    Lysates/proteins:25ug per lane.
    Blocking buffer:3%nonfat dry milk in TBST.
    Detection:ECL Basic Kit(RM00020).
    Exposure time:3s.
HRP偶联羊抗鼠二抗文献引用 : 1058

AS003

HRP-conjugated Goat anti-Mouse IgG (H+L)

  • 应用：WB,DB,ELISA
  • 物种：Mouse
  • Western blot analysis of lysates from HeLa cells,using GAPDH antibody as the primary antibody.
    Secondary antibody:HRP Goat Anti-Mouse IgG (H+L)antibody (AS003)at 1:5000/1:10000 dilution.
    Lysates/proteins:25ug per lane.
    Blocking buffer:3%nonfat dry milk in TBST.
    Detection:ECL Basic Kit(RM00020).
    Exposure time:60s.
HRP偶联羊抗鼠重链二抗文献引用 : 41

AS064

HRP-conjugated Goat anti-Mouse IgG Heavy Chain

  • 应用：WB
  • 物种：Mouse
  • Western blot analysis of lysates from Mouse and Rabbit IgG,using HRP-conjugated Goat Anti-Mouse IgG Heavy Chain
    (AS064)at 1:5000-1:40000 dilution.Lysates/proteins:100 ng per lane.Blocking buffer:3%nonfat dry milk in TBST.
    Detection:ECL Basic Kit(RM00020).Exposure time:10s.
二抗精品抗体
常见二抗
货号名称来源应用文献数量
AS071Goat Anti-Mouse IgG (H+L)GoatWB,ELISA,CUT&Tag13
AS070Goat Anti-Rabbit IgG(H+L)GoatWB,ELISA,CUT&Tag17
AS031HRP-conjugated Donkey anti-Goat IgG (H+L)DonkeyWB,IHC-P11
AS038HRP-conjugated Donkey anti-Rabbit IgG (H+L)DonkeyWB,IHC-P19
AS002HRP-conjugated Goat anti-Human IgG (H+L)GoatWB24
AS003HRP-conjugated Goat anti-Mouse IgG (H+L)GoatWB,DB,ELISA1058
AS064HRP-conjugated Goat anti-Mouse IgG Heavy ChainGoatWB41
AS014HRP-conjugated Goat anti-Rabbit IgG (H+L)GoatWB,DB,IHC-P,ELISA1998
AS028HRP-conjugated Goat anti-Rat IgG (H+L)GoatWB,IHC-P25
AS061HRP-conjugated Mouse anti-Rabbit IgG Light ChainMouseWB,IP63
AS029HRP-conjugated Rabbit anti-Goat IgG (H+L)RabbitWB,IHC-P59

试剂

实验流程产品名称备注货号
抗体稀释Western Blot Primary Antibody Dilution Buffer｛WB一抗稀释液｝为抗体提供稳定的工作环境，维持抗体活性与稳定性，减少非特异性结合RM02954
抗体稀释Universal Antibody Dilution Buffer｛通用型抗体稀释液｝兼容性强，可用于WB IHC IF的一抗和二抗的稀释RM02955
封闭Rapid Blocking Buffer｛快速封闭液(5*)｝兼容WB IHC IF的封闭RM02956
封闭Skimmed milk powder {脱脂奶粉}可用于WB封闭及抗体稀释RM00014

这是WB实验的最终信号检测与结果分析阶段。当带有HRP标记的二抗与目标蛋白结合后，加入化学发光底物。HRP酶会催化底物发生氧化反应并产生光信号。通过将膜置于化学发光成像仪下，即可捕获这些光信号，形成可见的条带图像。条带的位置对应于目标蛋白的分子量，而条带的亮度或强度则在理想条件下与目标蛋白的含量成正比。最后，通过专业的图像分析软件对条带进行定量分析，从而对目标蛋白在不同样本中的表达水平进行比较和评估。

试剂

实验流程产品名称规格货号
化学发光ECL Basic Plus Kit {ECL化学发光底物Plus(普通型)}检测灵敏度可达到10pg（皮克） HRPRM00020P
化学发光ECL Enhanced Plus Kit {ECL化学发光底物Plus(增强型)}检测灵敏度可达到pg（皮克） 至高 fg（飞克） HRPRM00021P
化学发光ECL Super Kit {ECL化学发光底物(超敏型)}检测灵敏度可达到更低 fg（飞克） HRPRM02867

仪器设备

化学发光成像系统

  • 高分辨率，4600w像素科学级CMOS（sCMOS）
  • 一键成像、一键多张保存、一键导入
  • 多种曝光模式，兼容不同丰度蛋白拍摄，减少调条件摸索

Western Blot (WB) 技术资源

WB实验技术指南

WB实验常见问题排查与应对指南

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