Recombinant
RabMAb

Recombinant Anti-CD22 antibody [EPR20061] (ab207727)

Overview

  • Product name

    Anti-CD22 antibody [EPR20061]
    See all CD22 primary antibodies
  • Description

    Rabbit monoclonal [EPR20061] to CD22
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, IPmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant fragment within Human CD22 aa 1-250. The exact sequence is proprietary.
    Database link: P20273

  • Positive control

    • WB: Ramos, Daudi and Raji whole cell lysates; Human tonsil and fetal spleen lysates. IHC-P: Human spleen and tonsil tissues. ICC/IF: Daudi and Raji cells. IP: Human tonsil lysate; Raji whole cell lysate.
  • 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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab207727 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 130 kDa (predicted molecular weight: 95 kDa).
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/100.
IP 1/30.

Target

  • Function

    Mediates B-cell B-cell interactions. May be involved in the localization of B-cells in lymphoid tissues. Binds sialylated glycoproteins; one of which is CD45. Preferentially binds to alpha-2,6-linked sialic acid. The sialic acid recognition site can be masked by cis interactions with sialic acids on the same cell surface. Upon ligand induced tyrosine phosphorylation in the immune response seems to be involved in regulation of B-cell antigen receptor signaling. Plays a role in positive regulation through interaction with Src family tyrosine kinases and may also act as an inhibitory receptor by recruiting cytoplasmic phosphatases via their SH2 domains that block signal transduction through dephosphorylation of signaling molecules.
  • Tissue specificity

    B-lymphocytes.
  • Sequence similarities

    Belongs to the immunoglobulin superfamily. SIGLEC (sialic acid binding Ig-like lectin) family.
    Contains 6 Ig-like C2-type (immunoglobulin-like) domains.
    Contains 1 Ig-like V-type (immunoglobulin-like) domain.
  • Domain

    Contains 4 copies of a cytoplasmic motif that is referred to as the immunoreceptor tyrosine-based inhibitor motif (ITIM). This motif is involved in modulation of cellular responses. The phosphorylated ITIM motif can bind the SH2 domain of several SH2-containing phosphatases.
  • Post-translational
    modifications

    Phosphorylation of Tyr-762, Tyr-807 and Tyr-822 are involved in binding to SYK, GRB2 and SYK, respectively. Phosphorylation of Tyr-842 is involved in binding to SYK, PLCG2 and PIK3R1/PIK3R2.
    Phosphorylated on tyrosine residues by LYN.
  • Cellular localization

    Cell membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • B cell receptor CD22 precursor antibody
    • B lymphocyte cell adhesion molecule antibody
    • B-cell receptor CD22 antibody
    • B-lymphocyte cell adhesion molecule antibody
    • BL CAM antibody
    • BL-CAM antibody
    • BLCAM antibody
    • CD 22 antibody
    • CD22 antibody
    • CD22 antigen antibody
    • CD22 molecule antibody
    • CD22 protein antibody
    • CD22_HUMAN antibody
    • Lectin 2 antibody
    • Leu14 antibody
    • Lyb8 antibody
    • MGC130020 antibody
    • sialic acid binding Ig like lectin 2 antibody
    • Sialic acid binding immunoglobulin like lectin 2 antibody
    • Sialic acid-binding Ig-like lectin 2 antibody
    • SIGLEC 2 antibody
    • Siglec-2 antibody
    • SIGLEC2 antibody
    • T cell surface antigen Leu 14 antibody
    • T-cell surface antigen Leu-14 antibody
    see all

Images

  • Immunohistochemical analysis of paraffin-embedded human spleen tissue labeling CD22 with ab207727 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Membranous and cytoplasmic staining on B cells of human spleen [PMID: 11967115]. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunofluorescent analysis of 100% methanol-fixed Daudi (Human Burkitt's lymphoma cell line) cells labeling CD22 with ab207727 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on Daudi cell line.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

  • Lanes 1 & 4 : Anti-CD22 antibody [EPR20061] (ab207727) at 1/2000 dilution
    Lanes 2-3 : Anti-CD22 antibody [EPR20061] (ab207727) at 1/10000 dilution

    Lane 1 : Ramos (Human Burkitt's lymphoma cell line) whole cell lysate
    Lane 2 : Daudi (Human Burkitt's lymphoma cell line) whole cell lysate
    Lane 3 : Raji (Human Burkitt's lymphoma cell line) whole cell lysate
    Lane 4 : Human tonsil 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: 95 kDa
    Observed band size: 130 kDa
    why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure times: Lane 1: 3 seconds; Lane 2/3: 1 second; Lane 4: 8 seconds.

    The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 24264377 & 21178016).

  • CD22 was immunoprecipitated from 0.35 mg of Human tonsil lysate with ab207727 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab207727 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

    Lane 1: Human tonsil lysate, 10 μg (Input).

    Lane 2: ab207727 IP in Human tonsil lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab207727 in Human tonsil lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 10 seconds.

  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling CD22 with ab207727 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Membranous and cytoplasmic staining on B cells of human tonsil [PMID: 11967115]. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunofluorescent analysis of 100% methanol-fixed Raji (Human Burkitt's lymphoma cell line) cells labeling CD22 with ab207727 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on Raji cell line.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

  • Anti-CD22 antibody [EPR20061] (ab207727) at 1/1000 dilution + Human fetal spleen lysate at 10 µg

    Secondary
    VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution

    Predicted band size: 95 kDa
    Observed band size: 130 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 24264377 & 21178016).

  • CD22 was immunoprecipitated from 0.35 mg of Raji (Human Burkitt's lymphoma cell line) whole cell lysate with ab207727 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab207727 at 1/2000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

    Lane 1: Raji whole cell lysate, 10 μg (Input).

    Lane 2: ab207727 IP in Raji whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab207727 in Raji whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 10 seconds.

  • Immunohistochemical analysis of paraffin-embedded human thymoma tissue labeling CD22 with ab207727 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Negative control: no staining on human thymoma [PMID: 11967115].

    Counter stained with Hematoxylin.

     

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

References

ab207727 has not yet been referenced specifically in any publications.

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