重组Anti-CD19抗体[EPR23174-145] (ab245235)
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD19 antibody [EPR23174-145] (ab245235)](https://www.abcam.cn/ps/products/245/ab245235/Images/ab245235-12-Anti-CD19-antibody-EPR23174-145-for-Immunohistochemistry-spleen-mouse.jpg "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD19 antibody [EPR23174-145] (ab245235)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23174-145] to CD19
- Suitable for: WB, Flow Cyt, IP, IHC-Fr, IHC-P, ICC/IF, mIHC
- Reacts with: Mouse
Related conjugates and formulations
概述
-
产品名称
Anti-CD19抗体[EPR23174-145]
参阅全部 CD19 一抗 -
描述
兔单克隆抗体[EPR23174-145] to CD19 -
宿主
Rabbit -
经测试应用
适用于: WB, Flow Cyt, IP, IHC-Fr, IHC-P, ICC/IF, mIHCmore details -
种属反应性
与反应: Mouse -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
-
阳性对照
- WB: Mouse spleen tissue lysate. WEHI-231 and A20 whole cell lysate. IHC-P: Mouse spleen tissue. ICC/IF: Mouse peripheral blood mononuclear cells. Flow Cyt: Mouse peripheral blood mononuclear cells. IP: Mouse spleen lysate. IHC-Fr: Mouse spleen tissue. mIHC: Mouse spleen tissue, mouse bone marrow, mouse thymus, mouse lymph node
-
常规说明
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
性能
-
形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading... -
纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR23174-145 -
同种型
IgG -
研究领域
相关产品
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab245235于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。
| 应用 | Ab评论 | 说明 |
|---|---|---|
| WB |
1/1000. Detects a band of approximately 60-120 kDa (predicted molecular weight: 60 kDa).
|
|
| Flow Cyt |
1/500.
|
|
| IP |
1/30.
|
|
| IHC-Fr | (1) |
1/250.
|
| IHC-P | (4) |
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
|
| ICC/IF |
1/50.
|
|
| mIHC |
1/1000.
|
| 说明 |
|---|
|
WB
1/1000. Detects a band of approximately 60-120 kDa (predicted molecular weight: 60 kDa). |
|
Flow Cyt
1/500. |
|
IP
1/30. |
|
IHC-Fr
1/250. |
|
IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
|
ICC/IF
1/50. |
|
mIHC
1/1000. |
靶标
-
功能
Assembles with the antigen receptor of B lymphocytes in order to decrease the threshold for antigen receptor-dependent stimulation. -
疾病相关
Defects in CD19 are the cause of immunodeficiency common variable type 3 (CVID3) [MIM:613493]; also called antibody deficiency due to CD19 defect. CVID3 is a primary immunodeficiency characterized by antibody deficiency, hypogammaglobulinemia, recurrent bacterial infections and an inability to mount an antibody response to antigen. The defect results from a failure of B-cell differentiation and impaired secretion of immunoglobulins; the numbers of circulating B cells is usually in the normal range, but can be low. -
序列相似性
Contains 2 Ig-like C2-type (immunoglobulin-like) domains. -
翻译后修饰
Phosphorylated on serine and threonine upon DNA damage, probably by ATM or ATR. Phosphorylated on tyrosine following B-cell activation. -
细胞定位
Membrane. - Information by UniProt
-
数据库链接
- Entrez Gene: 12478 Mouse
- SwissProt: P25918 Mouse
- Unigene: 4360 Mouse
-
别名
- Antibody deficiency due to defect in CD19 antibody
- Antibody deficiency due to defect in CD19, included antibody
- AW495831 antibody
see all
图片
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD19 antibody [EPR23174-145] (ab245235)This image is courtesy of an anonymous AbreviewImmunohistochemistry analysis (Formalin/PFA-fixed paraffin-embedded sections) of paraformaldehyde fixed mouse spleen tissue. Stained with ab245235 at 1/1000 dilution. Secondary antibody used was Dako EnVision+ System- HRP Labelled Polymer Anti-R. Blocking was done with 5% serum for 1 hour at 21°C. The sample was incubated with the primary antibody and 5% Normal Goat Serum for 19 hours at 4°C. Antigen retrieval method was heat mediated Tris/EDTA pH 9.0.
-
![Immunohistochemistry (Frozen sections) - Anti-CD19 antibody [EPR23174-145] (ab245235)](https://www.abcam.cn/ps/products/245/ab245235/Images/ab245235-11-Anti-CD19-antibody-EPR23174-145-for-Immunohistochemistry-spleen-tissue-mouse.jpg)
Immunohistochemistry (Frozen sections) - Anti-CD19 antibody [EPR23174-145] (ab245235)This image is courtesy of an anonymous AbreviewImmunohistochemistry (Frozen sections) analysis of acetone-fixed mouse spleen tissue staining with ab245235 at 1/1000 dilution. Secondary antibody was Dako EnVision+ System- HRP Labelled Polymer Anti-R Samples were incubated with the primary antibody with 5% Normal Goat Serum for 19 hours at 4°C. Blocking was done using 5% serum for 1 hour at 21°C -
![Multiplex immunohistochemistry - Anti-CD19 antibody [EPR23174-145] (ab245235)](https://www.abcam.cn/ps/products/245/ab245235/Images/ab245235-536455-anti-cd11b-cd19-ly6c-multiplex-immunohistochemsitry-spleen-mouse.JPG)
Multiplex immunohistochemistry - Anti-CD19 antibody [EPR23174-145] (ab245235)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse spleen labelling LY6C with ab317272 at 1/100 (B), CD11b with ab133357 at 1/20000 dilution (C) and CD19 with ab245235 at 1/1000 dilution (D). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Panel A: merged staining of anti-LY6C (green; Opal™690), anti-CD11b (magenta; Opal™570) and anti-CD19 (yellow; Opal™520) on mouse spleen.
Panel B: anti-LY6C stained on monocytes/macrophages.
Panel C: anti-CD11b stained on monocytes/macrophages.
Panel D: anti-CD19 stained on B cells.
Co-staining of LY6C and CD11b can be observed.The section was incubated in three rounds of staining: in the order of ab317272, ab133357, and ab245235 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope. -
![Immunohistochemistry (Frozen sections) - Anti-CD19 antibody [EPR23174-145] (ab245235)](https://www.abcam.cn/ps/products/245/ab245235/Images/ab245235-455278-AntiCD19-antibody-Immunocytochemistry.jpg)
Immunohistochemistry (Frozen sections) - Anti-CD19 antibody [EPR23174-145] (ab245235)Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized perfusion fixed frozen Mouse spleen tissue labeling CD19 with ab245235 at 1/250 (1.82 ug/ml) dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (Green). Positive staining on mouse spleen. is observed. The nuclear counterstain was DAPI (Blue).
Secondary antibody control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1000 dilution.
Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
-
![Western blot - Anti-CD19 antibody [EPR23174-145] (ab245235)](https://www.abcam.cn/ps/products/245/ab245235/Images/ab245235-6-anti-cd19-antibody-epr23174-145-western-blot-mouse.jpg)
Western blot - Anti-CD19 antibody [EPR23174-145] (ab245235)All lanes : Anti-CD19 antibody [EPR23174-145] (ab245235) at 1/1000 dilution
Lane 1 : WEHI-231 (mouse B cell lymphoma B lymphocyte), whole cell lysate
Lane 2 : A20 (mouse reticulum sarcoma B lymphocyte), whole cell lysate
Lane 3 : CTLL-2 (mouse T lymphocyte), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 60 kDa
Observed band size: 60-120 kDa why is the actual band size different from the predicted?Blocking and dilution buffer: 5% NFDM/TBS.
Exposure time: 48 seconds.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 26516065).
Negative control: CTLL-2 (PMID: 1370948) is treated as a negative cell line according to the literature, that CD19 is a B lymphocyte cell-surface marker that is expressed early during pre-B-cell differentiation, not in T cells.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD19 antibody [EPR23174-145] (ab245235)](https://www.abcam.cn/ps/products/245/ab245235/Images/ab245235-4-anti-cd19-antibody-epr23174-145-immunohistochemistry-spleen-mouse.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD19 antibody [EPR23174-145] (ab245235)Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling CD19 with ab245235 at 1/1000 dilution (0.446 µg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer). Positive staining on mouse spleen (PMID: 18648532). The section was incubated with ab245235 for 10 mins at room temperature. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
-
![Immunocytochemistry/ Immunofluorescence - Anti-CD19 antibody [EPR23174-145] (ab245235)](https://www.abcam.cn/ps/products/245/ab245235/Images/ab245235-1-anti-cd19-antibody-epr23174-145-immunocytochemistry-pbmc-mouse.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-CD19 antibody [EPR23174-145] (ab245235)Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized mouse PBMC (mouse primary peripheral blood mononuclear cell) cells labeling CD19 with ab245235 at 1/50 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing membranous staining in some subsets of mouse PBMCs. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.
-
![Flow Cytometry - Anti-CD19 antibody [EPR23174-145] (ab245235)](https://www.abcam.cn/ps/products/245/ab245235/Images/ab245235-2-anti-cd19-antibody-epr23174-145-flowcytometry-pbmc-mouse.jpg)
Flow Cytometry - Anti-CD19 antibody [EPR23174-145] (ab245235)Flow cytometric analysis of mouse peripheral blood mononuclear cell (PBMC) cells labeling CD19 with ab245235 at 1/500 dilution (Right) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Left). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
Cells were stained with rabbit IgG (Left) or ab245235 (Right). Then stained with anti-CD3 conjugated to Alexa Fluor® 647. CD19 and CD3 are mutually exclusive expressed in mouse PBMC.
Gated on viable cells.
-
![Immunoprecipitation - Anti-CD19 antibody [EPR23174-145] (ab245235)](https://www.abcam.cn/ps/products/245/ab245235/Images/ab245235-3-anti-cd19-antibody-epr23174-145-immunoprecipitation-spleen-mouse.jpg)
Immunoprecipitation - Anti-CD19 antibody [EPR23174-145] (ab245235)CD19 was immunoprecipitated from 0.35 mg mouse spleen lysate with ab245235 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab245235 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.
Lane 1: Mouse spleen lysate 10µg
Lane 2: ab245235 IP in mouse spleen lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab245235 in mouse spleen lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 seconds.
-
![Western blot - Anti-CD19 antibody [EPR23174-145] (ab245235)](https://www.abcam.cn/ps/products/245/ab245235/Images/ab245235-5-anti-cd19-antibody-epr23174-145-western-blot-spleen-mouse.jpg)
Western blot - Anti-CD19 antibody [EPR23174-145] (ab245235)Anti-CD19 antibody [EPR23174-145] (ab245235) at 1/1000 dilution + Mouse spleen tissue lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 60 kDa
Observed band size: 60-120 kDa why is the actual band size different from the predicted?Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 48 seconds.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 26516065).
-
![Anti-CD19 antibody [EPR23174-145] (ab245235)](https://www.abcam.cn/ps/products/245/ab245235/Images/ab245235-7-benefits-of-recombinant-antibodies.png)
Anti-CD19 antibody [EPR23174-145] (ab245235)
实验方案
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
数据表及文件
-
SDS download
-
Datasheet download
Certificate of Compliance
文献 (21)
ab245235 被引用在 21 文献中.
- Azevedo PO et al. Differential requirement of neutralizing antibodies and T cells on protective immunity to SARS-CoV-2 variants of concern. NPJ Vaccines 8:15 (2023). PubMed: 36781862
- Zhang Y et al. Extracellular Vesicles Expressing CD19 Antigen Improve Expansion and Efficacy of CD19-Targeted CAR-T Cells. Int J Nanomedicine 18:49-63 (2023). PubMed: 36636644
- Ma Y et al. Impact of chemotherapeutic agents on liver microenvironment: oxaliplatin create a pro-metastatic landscape. J Exp Clin Cancer Res 42:237 (2023). PubMed: 37697332
- Deng G et al. Ibrutinib Inhibits BTK Signaling in Tumor-Infiltrated B Cells and Amplifies Antitumor Immunity by PD-1 Checkpoint Blockade for Metastatic Prostate Cancer. Cancers (Basel) 15:N/A (2023). PubMed: 37190284
- Su D et al. Spatiotemporal single-cell transcriptomic profiling reveals inflammatory cell states in a mouse model of diffuse alveolar damage. Exploration (Beijing) 3:20220171 (2023). PubMed: 37933384