Recombinant Anti-CD22 antibody [EPR20061] (ab207727)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR20061] to CD22
- Suitable for: WB, IHC-P, ICC/IF, IP
- Reacts with: Human
Related conjugates and formulations
Overview
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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. This information is proprietary to Abcam and/or its suppliers.
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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.
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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: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR20061 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Related Products
Applications
The Abpromise guarantee
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 |
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WB |
1/1000. Detects a band of approximately 130 kDa (predicted molecular weight: 95 kDa).
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IHC-P | (2) |
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
1/100.
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IP |
1/30.
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Notes |
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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
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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
modificationsPhosphorylation 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
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Database links
- Entrez Gene: 933 Human
- Omim: 107266 Human
- SwissProt: P20273 Human
- Unigene: 579691 Human
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Alternative names
- B cell receptor CD22 precursor antibody
- B lymphocyte cell adhesion molecule antibody
- B-cell receptor CD22 antibody
see all
Images
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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.
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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.
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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).
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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.
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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.
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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 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 24264377 & 21178016).
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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.
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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.
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 (0)
ab207727 has not yet been referenced specifically in any publications.