Recombinant Anti-CD19 antibody [EPR23174-145] (ab245235)
![Immunohistochemistry (Frozen sections) - Anti-CD19 antibody [EPR23174-145] (ab245235)](https://www.abcam.com/ps/products/245/ab245235/Images/ab245235-455278-AntiCD19-antibody-Immunocytochemistry.jpg "Immunohistochemistry (Frozen 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
- Reacts with: Mouse
Related conjugates and formulations
Overview
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Product name
Anti-CD19 antibody [EPR23174-145]
See all CD19 primary antibodies -
Description
Rabbit monoclonal [EPR23174-145] to CD19 -
Host species
Rabbit -
Tested applications
Suitable for: WB, Flow Cyt, IP, IHC-Fr, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Mouse -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- 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 tiissue.
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General notes
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.
Properties
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Form
Liquid -
Storage instructions
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. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR23174-145 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab245235 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB |
1/1000. Detects a band of approximately 60-120 kDa (predicted molecular weight: 60 kDa).
|
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| Flow Cyt |
1/500.
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| IP |
1/30.
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| IHC-Fr | (1) |
1/250.
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| 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.
|
| Notes |
|---|
|
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. |
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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. |
Target
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Function
Assembles with the antigen receptor of B lymphocytes in order to decrease the threshold for antigen receptor-dependent stimulation. -
Involvement in disease
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. -
Sequence similarities
Contains 2 Ig-like C2-type (immunoglobulin-like) domains. -
Post-translational
modificationsPhosphorylated on serine and threonine upon DNA damage, probably by ATM or ATR. Phosphorylated on tyrosine following B-cell activation. -
Cellular localization
Membrane. - Information by UniProt
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Database links
- Entrez Gene: 12478 Mouse
- SwissProt: P25918 Mouse
- Unigene: 4360 Mouse
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Alternative names
- Antibody deficiency due to defect in CD19 antibody
- Antibody deficiency due to defect in CD19, included antibody
- AW495831 antibody
see all
Images
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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).
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![Western blot - Anti-CD19 antibody [EPR23174-145] (ab245235)](https://www.abcam.com/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.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD19 antibody [EPR23174-145] (ab245235)](https://www.abcam.com/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.
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![Immunocytochemistry/ Immunofluorescence - Anti-CD19 antibody [EPR23174-145] (ab245235)](https://www.abcam.com/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.
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![Flow Cytometry - Anti-CD19 antibody [EPR23174-145] (ab245235)](https://www.abcam.com/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.
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![Immunoprecipitation - Anti-CD19 antibody [EPR23174-145] (ab245235)](https://www.abcam.com/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.
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![Western blot - Anti-CD19 antibody [EPR23174-145] (ab245235)](https://www.abcam.com/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.com/ps/products/245/ab245235/Images/ab245235-7-benefits-of-recombinant-antibodies.png)
Anti-CD19 antibody [EPR23174-145] (ab245235)
Protocols
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.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (4)
ab245235 has been referenced in 4 publications.
- Zhang Y et al. SARS-CoV-2 rapidly adapts in aged BALB/c mice and induces typical pneumonia. J Virol N/A:N/A (2021). PubMed: 33692211
- Backx E et al. MECOM permits pancreatic acinar cell dedifferentiation avoiding cell death under stress conditions. Cell Death Differ 28:2601-2615 (2021). PubMed: 33762742
- Ding F et al. microRNA-375 released from extracellular vesicles of bone marrow mesenchymal stem cells exerts anti-oncogenic effects against cervical cancer. Stem Cell Res Ther 11:455 (2020). PubMed: 33109266
- Song Z et al. SARS-CoV-2 Causes a Systemically Multiple Organs Damages and Dissemination in Hamsters. Front Microbiol 11:618891 (2020). PubMed: 33510731