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Products:Immunology >> Adaptive Immunity >> B Cells >> CD
Overview
Product name
Anti-CD45 antibody
See all CD45 products (76) ...
Description
Rabbit polyclonal to CD45
Tested applications
Flow Cyt, IHC-Fr, WB, IHC-P, ICC/IFmore details
Cross reactivity
Reacts with
Mouse, Human, Rhesus monkey
Predicted to work with
Rat
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 900 - 1000 of Human CD45.
.
Positive control
This antibody gave a positive signal in Jurakt whole cell lysate and Human spleen tissue sections.
Properties
Form
Liquid
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage buffer
Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration
Concentration information loading...
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
Research areas
Immunology >> Adaptive Immunity >> Regulatory T Cells
Stem Cells >> Hematopoietic Progenitors >> Myeloid >> Mast Cell Lineage
Stem Cells >> Hematopoietic Progenitors >> Myeloid >> Neutrophil Lineage
Stem Cells >> Hematopoietic Progenitors >> Myeloid >> Monocytic Lineage
Stem Cells >> Hematopoietic Progenitors >> Myeloid >> Dendritic Cell Lineage
Stem Cells >> Hematopoietic Progenitors >> Lymphoid >> NK Cell Lineage
Stem Cells >> Hematopoietic Progenitors >> Lymphoid >> T Lymphocytic Lineage
Stem Cells >> Hematopoietic Progenitors >> Lymphoid >> B Lymphocytic Lineage
Stem Cells >> Hematopoietic Progenitors >> Surface Molecules
Neuroscience >> Cell Type Marker >> Glia marker >> Microglia marker
Signal Transduction >> Protein Phosphorylation >> Tyrosine Phosphatases
Immunology >> Cell Type Markers >> CD >> Non-lineage
Immunology >> Adaptive Immunity >> T Cells >> CD
Immunology >> Adaptive Immunity >> B Cells >> CD
Applications
Show applications key
Our Abpromise guarantee covers the use of ab10558 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Flow Cyt: Use 1µg for 106 cells.
IHC-Fr: 1/150
WB: 1/500Detects a band of approximately 155 kDa (predicted molecular weight: 147 kDa).Can be blocked with CD45 peptide (ab17553).
IHC-P: Use a concentration of 1 µg/mlPerform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF: 1/100
Target
Function
Protein tyrosine-protein phosphatase required for T-cell activation through the antigen receptor. Acts as a positive regulator of T-cell coactivation upon binding to DPP4. The first PTPase domain has enzymatic activity, while the second one seems to affect the substrate specificity of the first one. Upon T-cell activation, recruits and dephosphorylates SKAP1 and FYN.
Involvement in disease
Defects in PTPRC are a cause of severe combined immunodeficiency autosomal recessive T-cell-negative/B-cell-positive/NK-cell-positive (T(-)B(+)NK(+) SCID) [MIM:608971]. A form of severe combined immunodeficiency (SCID), a genetically and clinically heterogeneous group of rare congenital disorders characterized by impairment of both humoral and cell-mediated immunity, leukopenia, and low or absent antibody levels. Patients present in infancy recurrent, persistent infections by opportunistic organisms. The common characteristic of all types of SCID is absence of T-cell-mediated cellular immunity due to a defect in T-cell development.
Genetic variations in PTPRC are involved in multiple sclerosis susceptibility (MS) [MIM:126200]. MS is a neurodegenerative disorder characterized by the gradual accumulation of focal plaques of demyelination particularly in the periventricular areas of the brain. Peripheral nerves are not affected. Onset usually in third or fourth decade with intermittent progression over an extended period. The cause is still uncertain.
Sequence similarities
Belongs to the protein-tyrosine phosphatase family. Receptor class 1/6 subfamily.
Contains 2 fibronectin type-III domains.
Contains 2 tyrosine-protein phosphatase domains.
Domain
The first PTPase domain interacts with SKAP1.
Post-translational
modifications
Heavily N- and O-glycosylated.
Cellular localization
Membrane. Membrane raft. Colocalized with DPP4 in membrane rafts.
Target information above from: UniProt accessionP08575
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Alternative names
- B220 antibody
- CD 45 antibody
- CD45 antibody
see all
Database links
- Entrez Gene: 5788 Human
- Entrez Gene: 19264 Mouse
- Entrez Gene: 24699 Rat
- Omim: 151460 Human
- SwissProt: P08575 Human
- SwissProt: P06800 Mouse
- SwissProt: P04157 Rat
- Unigene: 654514 Human
see all
Anti-CD45 antibody images:
Western blot - Anti-CD45 antibody (ab10558)
All lanes : Anti-CD45 antibody (ab10558) at 1/500 dilution
Lane 1 : Jurkat Whole Cell Lysate
Lane 2 : Jurkat Whole Cell Lysate with CD45 peptide (ab17553)
Lysates/proteins at 20 µg per lane.
Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP) (ab6721) at 1/5000 dilution
Predicted band size : 147 kDa
Exposure time : 3 minutes
Flow Cytometry - Anti-CD45 antibody (ab10558)
Asynchronous KM-H2 cells were pelleted and labeled by indirect immunofluorescence. Cells were stained with ab10558 (1/200) for 30min at 4'C, washed and then stained with goat anti-rabbit alexafluor 488 (1/200). Forward/Side scatter were used to eliminate cellular debris. The accompanying marker was applied such that only 2% of the IgG control was positive Based on the accompanying image, approximately 8.4% of cells exhibited positive staining for anti-CD45. Since KM-H2 are known to have low levels of CD45 transcripts they are expected to have low levels of CD45, which is reflected in the ~8%. This image is from an Abreview.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - CD45 antibody (ab10558)
ab10558 (1:40) staining CD45 in paraffin-embedded human tonsil (left panel) using an automated system (Ventana Discovery). Right-hand panel shows negative control (no primary antibody).
Using this protocol there is strong membrane staining of B cells in the germinal centres and mantle zone of the follicles and scattered cells of the interfollicular areas (paracortical T and B cells). There is a mild to moderate degree of cytoplasmic staining associated with the membrane staining in these specific cells.
Sections were rehydrated and antigen retrieved in CC1 Cell Conditioning Buffer using Ventana Extended Retrieval programme. Slides were blocked in 3% H2O2 /4 min/ 37°C and incubated with ab10558 (1:40 dilution / 1 hour/ 37°C). Sections then blocked (4mins/ 37°C) and incubated with Dako swine anti-rabbit antibody (1:50, 28 min/ 37°C). Staining was amplified and detected by incubation with Ventana Streptavidin ABC system (16 min/ 37°C) and Ventana DAB map reagent (8 min/ 37°C). Slides were counterstained with Haematoxylin and coverslipped in DPX.
For manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
Immunohistochemistry (Frozen sections) - CD45 antibody (ab10558)
ab10558 staining CD45 in fetal mouse liver tissue sections 12.5 dpc by Immunohistochemistry (frozen sections). Tissue was fixed with paraformaldehyde and a permeabilization step was performed using 0.1% Tween-20. Samples were then blocked with 5% BSA for 1.5 hours at 20°C followed by incubation with the primary antibody at a 1/150 dilution, for 16 hours at 4°C. A Goat anti-rabbit Alexa Fluor® 594 polyclonal was used as the secondary antibody at a 1/1500 diution.
Image courtesy of Mr Sam Lee by Abreview.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - CD45 antibody (ab10558)
IHC image of CD45 staining in human spleen formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab10558, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
Flow Cytometry - Anti-CD45 antibody (ab10558)
Overlay histogram showing Jurkat cells stained with ab10558 (red line). The cells were fixed with 80% methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab10558, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/1000 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.
References for Anti-CD45 antibody (ab10558)
This product has been referenced in:
- O'Hara Let al. Androgen receptor expression in the caput epididymal epithelium is essential for development of the initial segment and epididymal spermatozoa transit. Endocrinology 152:718-29 (2011). IHC-P; Mouse.Read more (PubMed: 21177831) »
- Biscetti Fet al. High-mobility group box-1 protein promotes angiogenesis after peripheral ischemia in diabetic mice through a VEGF-dependent mechanism. Diabetes 59:1496-505 (2010). IHC-P; Mouse.Read more (PubMed: 20200317) »
See all 5 publications for this product
Publishing research using ab10558? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"