Recombinant
RabMAb

Recombinant Anti-CD19 antibody [EPR5906] (ab134114)

Rabbit recombinant monoclonal CD19 antibody [EPR5906]. Validated in WB, IHC, Flow Cyt, ICC/IF and tested in Human. Cited in 8 publication(s).

Overview

  • Product name

    Anti-CD19 antibody [EPR5906]
    See all CD19 primary antibodies
  • Description

    Rabbit monoclonal [EPR5906] to CD19
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, Flow Cytmore details
    Unsuitable for: IP
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human CD19 aa 300 to the C-terminus. The exact sequence is proprietary.

  • Positive control

    • WB: Namalwa and Ramos cell lysates and human tonsil tissue lysate. IHC-P: Human tonsil, diffuse large B-cell lymphoma, B-cell chronic lymphocytic leukaemia and spleen tissue. ICC/IF: Raji cells. Flow Cyt: Raji cells.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab134114 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 - 1/10000. Detects a band of approximately 95 kDa (predicted molecular weight: 61 kDa).
IHC-P 1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

ICC/IF 1/500 - 1/1000.
Flow Cyt Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for IP.
  • 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.
    • Information by UniProt
    • Database links

    • Alternative names

      • 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

    • Flow cytometry analysis of Raji cells (Human Burkitt's lymphoma B lymphocyte) labelling CD19 with ab134114 at 1:1000 dilution, 1.186 µg/ml (red). Cells were fixed with 4% paraformaldehyde, permeabilised with 90% methanol. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/2000.

      Isotype control (black) - Rabbit monoclonal IgG (ab172730)

      Unlabeled control (blue) - Unlabelled cells

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling CD19 with unpurified ab134114 at a dilution of 1/250.

    • Immunocytochemistry/Immunofluorescence analysis of Raji cells labelling CD19 with purified ab134114 at a dilution of 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

      Control 1: primary antibody (1/500) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).

      Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).

    • Lanes 1-3 : Anti-CD19 antibody [EPR5906] (ab134114) at 1/10000 dilution
      Lane 4 : Anti-CD19 antibody [EPR5906] (ab134114) at 1/2000 dilution

      Lane 1 : NAMALWA (Human Burkitt's lymphoma) whole cell lysate)
      Lane 2 : Human tonsil tissue lysate
      Lane 3 : Jurkat (Human acute T cell leukemia)whole cell lysate
      Lane 4 : RAMOS (Human Burkitt's lymphoma) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 61 kDa
      Observed band size: 95 kDa
      why is the actual band size different from the predicted?



      Exposure time:
      Lane 1-3: 3 minutes
      Lane 4: 30 seconds

      Jurkat is a CD19 null cell line according to the paper (PMID:  19147785).

    • All lanes : Anti-CD19 antibody [EPR5906] (ab134114) at 1/5000 dilution (purified)

      Lane 1 : Namalwa whole cell lysate
      Lane 2 : Daudi whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size: 61 kDa
      Observed band size: 95 kDa why is the actual band size different from the predicted?



      Blocking and dilution buffer: 5% NFDM/TBST
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling CD19 with purified ab134114 at a dilution of 1/500. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    • Negative tissue: Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling CD19 with purified ab134114 at a dilution of 1/500. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human diffuse large B-cell lymphoma tissue labelling CD19 with unpurified ab134114.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human spleen tissue labelling CD19 with unpurified ab134114.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human B-cell chronic lymphocytic leukaemia tissue labelling CD19 with unpurified ab134114.

    • Immunohistochemical analysis of paraffin embedded human skeletal muscle tissue using unpurified ab134114 showing negative staining.

    References

    This product has been referenced in:

    • Matsuda M  et al. Human NK cell development in hIL-7 and hIL-15 knockin NOD/SCID/IL2rgKO mice. Life Sci Alliance 2:N/A (2019). Read more (PubMed: 30936185) »
    • Wang C  et al. Whole-genome sequencing reveals genomic signatures associated with the inflammatory microenvironments in Chinese NSCLC patients. Nat Commun 9:2054 (2018). Read more (PubMed: 29799009) »
    See all 9 Publications for this product

    Customer reviews and Q&As

    1-4 of 4 Abreviews or Q&A

    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (IM9 cells)
    Gel Running Conditions
    Reduced Denaturing (4-20% Tris Glycin Gel, semi dry transfer on PVDF membrane)
    Loading amount
    25 µg
    Specification
    IM9 cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

    Abcam user community

    Verified customer

    Submitted Mar 26 2019

    Application
    Flow Cytometry
    Sample
    Human Cell (IM9 cells)
    Permeabilization
    No
    Gating Strategy
    SSC:0.1-768 FSC:256-600
    Specification
    IM9 cells
    Preparation
    Cell harvesting/tissue preparation method: Cells in suspension spinned down
    Sample buffer: PBS with 0.5% BSA
    Fixation
    Formaldehyde

    Abcam user community

    Verified customer

    Submitted Mar 26 2019

    Answer

    We have many products in catalogue. Please find below few of these

    https://www.abcam.com/cd19-antibody-epr5906-ab134114.html

    https://www.abcam.com/cd19-antibody-epr5906-ab134114.html

    https://www.abcam.com/cd19-antibody-2e2b6b10-ab31947.html

    https://www.abcam.com/baff-antibody-buffy-2-ab16081.html

    https://www.abcam.com/baff-antibody-ab117256.html

    The compatible secondary antibodies can be found using the below link

    https://www.abcam.com/Search?AppliedFacets.FacetProductType=Secondary+antibodies

    We have complete set of IHC kits available. Please check the link;

    https://www.abcam.com/index.html?pageconfig=resource&rid=12906

    Read More

    Answer

    Thank you for your enquiry regarding ab134114.

    This is a monoclonal rabbit antibody and we do not provide the concentration for it. These types of antibodies have relatively low IgG concentration, however their affinity and specificity are very high. You may wish to read this page for further information:

    https://www.abcam.com/index.html?pageconfig=resource&rid=15279

    Instead of providing the concentration, we normally list the recommended working dilution factors for the tested applications:

    WB: 1/1000 - 1/10000

    IP: 1/10 - 1/100

    IHC-P: 1/250 - 1/500

    ICC: 1/500 - 1/1000

    If you need any further assistance in the future, please do not hesitate to contact me.

    Read More

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