Overview

  • Product name
  • Description
    Rabbit polyclonal to CD45
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, Flow Cyt, IHC-P, IHC-Fr, ICC/IFmore details
  • Species reactivity
    Reacts with: Human, Pig, Monkey
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1250 to the C-terminus of Human CD45.

    (Peptide available as ab17550.)

  • Positive control
    • This antibody gave a positive signal in Jurkat whole cell lysate and Hodgkins lymphoma tissue sections.

Properties

Applications

Our Abpromise guarantee covers the use of ab10559 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 1/500. Detects a band of approximately 155 kDa (predicted molecular weight: 147 kDa).
Flow Cyt Use at an assay dependent concentration.

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IHC-Fr Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.

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.
  • Information by UniProt
  • Database links
  • Alternative names
    • B220 antibody
    • CD 45 antibody
    • CD45 antibody
    • CD45 antigen antibody
    • CD45R antibody
    • GP180 antibody
    • L-CA antibody
    • LCA antibody
    • Leukocyte common antigen antibody
    • loc antibody
    • Ly-5 antibody
    • LY5 antibody
    • Ly5, homolog of antibody
    • Lyt-4 antibody
    • OTTHUMP00000033813 antibody
    • OTTHUMP00000033816 antibody
    • OTTHUMP00000033817 antibody
    • OTTHUMP00000038574 antibody
    • Protein tyrosine phosphatase receptor type c polypeptide antibody
    • Protein tyrosine phosphatase, receptor type C antibody
    • protein tyrosine phosphatase, receptor type, C antibody
    • Protein tyrosine phosphatase, receptor type, c polypeptide antibody
    • Ptprc antibody
    • PTPRC_HUMAN antibody
    • Receptor-type tyrosine-protein phosphatase C antibody
    • T200 antibody
    • T200 glycoprotein antibody
    • T200 leukocyte common antigen antibody
    see all

Images

  • IHC image of ab10559 staining in human Hodgkins lymphoma 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 ab10559, 5µg/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.
    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • ab10559  rabbit polyclonal to CD45  (1/500) with secondary goat anti-rabbit IgG ab6721 (1/5000).

    Expected molecular weight: 147 kDa. WB exposure time: 1min 30sec    

    Lanes 1 to 10: 20µg of cell lysate per lane
    Lanes 6 to 10: ab17550 CD45 blocking peptide used at 1µg/ml

    Lane 1: HeLa Nuclear Extract (ab10559)
    Lane 2: HeLa Whole Cell Lysate (ab10559)
    Lane 3: A431 Whole Cell Lysate (ab10559)
    Lane 4: Jurkat Whole Cell Lysate (ab10559)
    Lane 5: HEK293 Whole Cell Lysate (ab10559)
    Lane 6: HeLa Nuclear Extract (ab10559+ ab17550)
    Lane 7: HeLa Whole Cell Lysate (ab10559+ ab17550)
    Lane 8: A431 Whole Cell Lysate (ab10559+ ab17550)
    Lane 9: Jurkat Whole Cell Lysate (ab10559+ ab17550)
    Lane 10: HEK293 Whole Cell Lysate (ab10559+ ab17550)

    A strong band, slightly higher was seen in Jurkat cell lysate. The band was blocked using the immunising peptide (ab17550). It is likely that the band is CD45.

  • Asynchronous KM-H2 cells were pelleted and labeled by indirect immunofluorescence. Cells were stained with ab10559 (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 9.62% of cells exhibited positive staining for anti-CD45. Since KM-H2 have low levels of CD45 transcripts, it is expected that they have low levels of CD45 on their surface which is reflected in the ~9% positive. This image is taken from an Abreview.

References

This product has been referenced in:

See all 13 Publications for this product

Customer reviews and Q&As

Filter by Application

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Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Sample
Monkey Tissue sections (Spleen)
Specification
Spleen
Permeabilization
No
Fixative
Formaldehyde
Username

Mr. Carl Hobbs

Verified customer

Submitted Aug 11 2014

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Sample
Human Tissue sections (Tonsil)
Specification
Tonsil
Permeabilization
No
Fixative
Formaldehyde
Username

Mr. Carl Hobbs

Verified customer

Submitted Aug 10 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Pig Tissue sections (Epithelial)
Specification
Epithelial
Fixative
Acetone
Permeabilization
No
Username

Mr. Daniel Holod-Betz

Verified customer

Submitted Apr 11 2012

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (spleen)
Specification
spleen
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Dako Target Retreival Solution
Permeabilization
No
Blocking step
5 minutes of peroxidase block then 10 minutes of protein block. These are ready-to-use reagents purchased from Dako as blocking agent for 15 minute(s) · Concentration: 100% · Temperature: 20°C
Username

Mr. Antibody Solutions

Verified customer

Submitted Apr 07 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (tonsil)
Specification
tonsil
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Dako Target Retreival Solution
Permeabilization
No
Blocking step
5 minutes of peroxidase block then 10 minutes of protein block. These are ready-to-use reagents purchased from Dako as blocking agent for 15 minute(s) · Concentration: 100% · Temperature: 20°C
Username

Mr. Antibody Solutions

Verified customer

Submitted Mar 26 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Flow Cytometry
Sample
Human Cell (KM-H2)
Specification
KM-H2
Preparation
Cell harvesting/tissue preparation method: Asynchronous KM-H2 were harvested and labeled by indirect immunofluorescence
Sample buffer: PBS + 5% FBS
Permeabilization
No
Gating Strategy
Forward/Side scatter were used to eliminate cellular debris. The accompanying marker was applied such that only 2% of the IgG control was positive.
Username

Dr. Kirk Mcmanus

Verified customer

Submitted Dec 16 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (PBMC)
Permeabilization
Yes - Triton X
Specification
PBMC
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Oct 09 2007

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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