Western blot analysis of extracts of Human serum, using CD19 antibody (A19013) at 1:1000 dilution.
Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Enhanced Kit (RM00021).
Exposure time: 3min.
Immunohistochemistry analysis of paraffin-embedded Human B-cell lymphoma（200X） using CD19 Rabbit mAb (A19013) at dilution of 1:200 (40x lens).Perform high pressure antigen retrieval with 10 mM citrate buffer pH 6.0 before commencing with IHC staining protocol.
Immunohistochemistry analysis of paraffin-embedded Human B-cell lymphoma（400X） using CD19 Rabbit mAb (A19013) at dilution of 1:200 (40x lens).Perform high pressure antigen retrieval with 10 mM citrate buffer pH 6.0 before commencing with IHC staining protocol.
Immunohistochemistry analysis of paraffin-embedded Human tonsil（200X） using CD19 Rabbit mAb (A19013) at dilution of 1:200 (40x lens).Perform high pressure antigen retrieval with 10 mM citrate buffer pH 6.0 before commencing with IHC staining protocol.
Immunohistochemistry analysis of paraffin-embedded human tonsil using CD19 Rabbit mAb (A19013) at dilution of 1:200 (40x lens).Perform high pressure antigen retrieval with 10 mM citrate buffer pH 6.0 before commencing with IHC staining protocol.
| 反应物种 | Human |
|---|---|
| 同种型 | IgG |
| 克隆号 | ARC0418 |
| 产品应用 | 测试种属及综合评分 |
| IHC-P | Human: |
| WB | Human: |
| 推荐稀释比 |
|
| 浓度查询 | 请输入产品标签上的lot号，例如4000000001 |
| 理论分子量 | 61kDa |
| 实际分子量 | 95kDa |
| 产品形式 | Liquid |
| 存储缓冲液 | Store at -20℃. Avoid freeze / thaw cycles. Buffer: PBS with 0.05% proclin300,0.05% BSA,50% glycerol,pH7.3. |
| 偶联物 | Unconjugated |
| 阳性样本 | Human serum |
| 细胞定位 | Membrane,Single-pass type I membrane protein |
| 纯化方式 | Affinity purification |
| 免疫原 | A synthetic peptide corresponding to a sequence within amino acids 457-556 of human CD19 (NP_001761.3). |
|---|---|
| 序列 | NAESYENEDEELTQPVARTMDFLSPHGSAWDPSREATSLGSQSYEDMRGILYAAPQLRSIRGQPGPNHEEDADSYENMDNPDGPDPAWGGGGRMGTWSTR |
一、实验试剂及耗材
二、实验步骤
（1）细胞收集
（2）总蛋白提取
（3）蛋白浓度测定（BCA法）
（1）制胶器安装
（2）分离胶制备
（3）浓缩胶制备
（4）上样
（5）电泳
（1）准备工作
（2）转膜
附表1：BSA标准品稀释
| 1mg/ml BSA(μl) | 稀释液(μl) | 对应的蛋白含量（mg/ml） |
|---|---|---|
| 0 | 20 | 0 |
| 1 | 19 | 0.05 |
| 2 | 18 | 0.1 |
| 4 | 16 | 0.2 |
| 8 | 12 | 0.4 |
| 12 | 8 | 0.6 |
| 16 | 4 | 0.8 |
| 20 | 0 | 1 |
附表2：分离胶浓度的选择
| 分子量（kDa) | 分离胶浓度 |
|---|---|
| X≤10 | 15% |
| 10<X≤15 | 13.5% |
| 15<X≤25 | 12% |
| 25<X≤35 | 11% |
| 35<X≤40 | 10% |
| 40<X≤55 | 9% |
| 55<X≤70 | 8% |
| 70<X≤100 | 7% |
| 100<X | 6% |
附表3：分离胶配制
| 分离胶浓度 | 6% | 7% | 8% | 9% | 10% | 11% | 12% | 13.5% | 15% |
|---|---|---|---|---|---|---|---|---|---|
| 去离子水（mL） | 5.3 | 4.9 | 4.6 | 4.3 | 4.0 | 3.65 | 3.3 | 2.8 | 2.3 |
| 30%丙烯酰胺 （mL） | 2 | 2.4 | 2.7 | 3.0 | 3.3 | 3.65 | 4.0 | 4.5 | 5.0 |
| 1.5M Tris-HCL(pH8.8) （mL） | 2.5 | ||||||||
| 10%SDS（mL） | 0.1 | ||||||||
| 10%AP（mL） | 0.1 | ||||||||
| TEMED（mL） | 0.01 | ||||||||
| 总体积（mL） | 10 |
附表4：浓缩胶配制
| 5%浓缩胶 | 5%浓缩胶 | 5%浓缩胶 | 5%浓缩胶 | 5%浓缩胶 | |
|---|---|---|---|---|---|
| 去离子水（mL） | 1.1 | 2.1 | 2.7 | 3.4 | 4.1 |
| 30%丙烯酰胺 （mL） | 0.33 | 0.5 | 0.67 | 0.83 | 1.0 |
| 1.0M Tris-HCL(pH6.8) （mL） | 0.25 | 0.38 | 0.5 | 0.63 | 0.75 |
| 10%SDS（mL） | 0.02 | 0.03 | 0.04 | 0.05 | 0.06 |
| 10%AP（mL） | 0.02 | 0.03 | 0.04 | 0.05 | 0.06 |
| TEMED（mL） | 0.002 | 0.003 | 0.004 | 0.005 | 0.006 |
| 总体积（mL） | 2 | 3 | 4 | 5 | 6 |
附表5：转膜条件选择
| 分子量（kDa) | 转膜条件 |
|---|---|
| X≤10 | 恒流200mA,30min |
| 10 < X≤15 | 恒流200mA,40min |
| 15< X≤20 | 恒流200mA,50min |
| 20< X≤50 | 恒流200mA, 1kDa/min+20min |
| 50< X≤100 | 恒流200mA, 1kDa/min+30min |
| 100< X≤150 | 恒流250mA, 1kDa/min+20min |
| 150< X≤180 | 恒流270mA, 1kDa/min(时间控制在3h内) |
| 180< X | 恒流270mA, 1kDa/min(时间控制在4h内) |
一、实验仪器及试剂
微波炉、4℃冰箱、恒温恒湿箱、烘箱、光学显微镜、移液器、孵育湿盒、抗原修复盒。
| 试剂名称 | 1*PBS/L | 试剂名称 | 1*PBST/L |
| NaH2PO4 | 0.23g | 1*PBS溶液 | 1L |
| Na2HPO4 | 1.15g | Tween 20 溶液 | 0.001L |
| NaCl | 8.5g | 用稀HCl溶液和NaOH溶液调pH值 | |
二、实验步骤
This gene encodes a member of the immunoglobulin gene superfamily. Expression of this cell surface protein is restricted to B cell lymphocytes. This protein is a reliable marker for pre-B cells but its expression diminishes during terminal B cell differentiation in antibody secreting plasma cells. The protein has two N-terminal extracellular Ig-like domains separated by a non-Ig-like domain, a hydrophobic transmembrane domain, and a large C-terminal cytoplasmic domain. This protein forms a complex with several membrane proteins including complement receptor type 2 (CD21) and tetraspanin (CD81) and this complex reduces the threshold for antigen-initiated B cell activation. Activation of this B-cell antigen receptor complex activates the phosphatidylinositol 3-kinase signalling pathway and the subsequent release of intracellular stores of calcium ions. This protein is a target of chimeric antigen receptor (CAR) T-cells used in the treatment of lymphoblastic leukemia. Mutations in this gene are associated with the disease common variable immunodeficiency 3 (CVID3) which results in a failure of B-cell differentiation and impaired secretion of immunoglobulins. CVID3 is characterized by hypogammaglobulinemia, an inability to mount an antibody response to antigen, and recurrent bacterial infections. Alternative splicing results in multiple transcript variants encoding distinct isoforms.
关联靶点
关联领域
.* 该数据来源于赛特新思(citexs.com)
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