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  1. Link

    cd19-antibody-epr23174-145-ab245235.pdf

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Immunology Adaptive Immunity B Cells CD
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RecombinantRabMAb

Recombinant Anti-CD19 antibody [EPR23174-145] (ab245235)

  • Datasheet
  • SDS
  • Certificate of Compliance
Reviews (5) Submit a question References (4)

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Immunohistochemistry (Frozen sections) - Anti-CD19 antibody [EPR23174-145] (ab245235)
  • Western blot - Anti-CD19 antibody [EPR23174-145] (ab245235)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD19 antibody [EPR23174-145] (ab245235)
  • Immunocytochemistry/ Immunofluorescence - Anti-CD19 antibody [EPR23174-145] (ab245235)
  • Flow Cytometry - Anti-CD19 antibody [EPR23174-145] (ab245235)
  • Immunoprecipitation - Anti-CD19 antibody [EPR23174-145] (ab245235)
  • Western blot - Anti-CD19 antibody [EPR23174-145] (ab245235)
  • 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

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Overview

  • 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.

  • 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.
  • 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
    For more information see here.

    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

  • 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

    • Immunology
    • Adaptive Immunity
    • B Cells
    • CD
    • Stem Cells
    • Hematopoietic Progenitors
    • Hematopoietic Stem Cells
    • Human Lineage Negative
    • Stem Cells
    • Hematopoietic Progenitors
    • Lymphoid
    • B Lymphocytic Lineage

Associated products

  • Alternative Versions

    • Anti-CD19 antibody [EPR23174-145] - BSA and Azide free (ab267394)
  • Compatible Secondaries

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)
    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

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).
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.
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.
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

  • 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
    modifications

    Phosphorylated on serine and threonine upon DNA damage, probably by ATM or ATR. Phosphorylated on tyrosine following B-cell activation.
  • Cellular localization

    Membrane.
  • Target information above from: UniProt accession P15391 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 12478 Mouse
    • SwissProt: P25918 Mouse
    • Unigene: 4360 Mouse
    • Alternative names

      • Antibody deficiency due to defect in CD19 antibody
      • Antibody deficiency due to defect in CD19, included antibody
      • AW495831 antibody
      • B lymphocyte antigen CD19 antibody
      • B lymphocyte surface antigen B4 antibody
      • B-lymphocyte antigen CD19 antibody
      • B-lymphocyte surface antigen B4 antibody
      • B4 antibody
      • CD19 antibody
      • CD19 antigen antibody
      • CD19 molecule antibody
      • Cd19 protein antibody
      • CD19_HUMAN antibody
      • CVID3 antibody
      • Differentiation antigen CD19 antibody
      • Leu 12 antibody
      • Leu-12 antibody
      • Leu12 antibody
      • MGC109570 antibody
      • MGC12802 antibody
      • T-cell surface antigen Leu-12 antibody
      see all

    Images

    • Immunohistochemistry (Frozen sections) - Anti-CD19 antibody [EPR23174-145] (ab245235)
      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)
      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)
      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)
      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)
      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)
      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)
      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)
      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.

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (4)

    Publishing research using ab245235? Please let us know so that we can cite the reference in this datasheet.

    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

    Customer reviews and Q&As

    Show All Reviews Q&A
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    1-5 of 5 Abreviews or Q&A

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD19 antibody [EPR23174-145]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (BL6 mouse lungs)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: pH6 30min steamer
    Permeabilization
    No
    Specification
    BL6 mouse lungs
    Blocking step
    Zytomed Systems HRP Polymer Protein Block as blocking agent for 5 minute(s) · Concentration: 100% · Temperature: 20°C
    Fixative
    Paraformaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Nov 26 2021

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD19 antibody [EPR23174-145]

    Inconclusive
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (palatine tonsil)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9.0
    Permeabilization
    No
    Specification
    palatine tonsil
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Fixative
    Paraformaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Jan 11 2021

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD19 antibody [EPR23174-145]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (spleen)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9.0
    Permeabilization
    No
    Specification
    spleen
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Fixative
    Paraformaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Jan 11 2021

    Immunohistochemistry (Frozen sections) abreview for Anti-CD19 antibody [EPR23174-145]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (Spleen)
    Permeabilization
    No
    Specification
    Spleen
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Fixative
    Acetone
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Jan 11 2021

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD19 antibody [EPR23174-145]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (tumor)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: FLEX taget retrival solution high pH (Dako)
    Permeabilization
    No
    Specification
    tumor
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 22°C
    Fixative
    Paraformaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Jun 25 2020

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