Key features and details
- Rabbit polyclonal to CD45
- Suitable for: Flow Cyt, WB, IHC-P
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
CD45 一次抗体 製品一覧
製品の詳細Rabbit polyclonal to CD45
アプリケーション適用あり: Flow Cyt, WB, IHC-Pmore details
種交差性交差種: Mouse, Rat, Human
交差が予測される動物種: Pig, Rhesus monkey
Synthetic peptide corresponding to Human CD45 aa 900-1000 conjugated to keyhole limpet haemocyanin.
(Peptide available as
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Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
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保存方法Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Preservative: 0.02% Sodium azide
Constituents: PBS, 1% BSA
Concentration information loading...
精製度Immunogen affinity purified
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.
ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.
|WB||1/500. Detects a band of approximately 190 kDa (predicted molecular weight: 147 kDa).|
|IHC-P||Use a concentration of 0.5 - 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
機能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.
関連疾患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.
配列類似性Belongs to the protein-tyrosine phosphatase family. Receptor class 1/6 subfamily.
Contains 2 fibronectin type-III domains.
Contains 2 tyrosine-protein phosphatase domains.
ドメインThe first PTPase domain interacts with SKAP1.
翻訳後修飾Heavily N- and O-glycosylated.
細胞内局在Membrane. Membrane raft. Colocalized with DPP4 in membrane rafts.
- Information by UniProt
- B220 antibody
- CD 45 antibody
- CD45 antibody
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 (HRP-DAB) system (16 min/ 37°C) before being counterstained with hematoxylin.
ab10558 staining CD45 in Human tonsil tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Negative control is shown in panel. Blocking was with horse serum (1/75) for 1 hour at room temperature. Samples were incubated with primary antibody (1/10) overnight at 4°C. A Biotin-conjugated Horse anti-mouse polyclonal (1/200) was used as the secondary antibody.
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.
All lanes : Anti-CD45 antibody (ab10558) at 1/500 dilution
Lane 1 : Jurkat Whole Cell Lysate
Lane 2 : Jurkat Whole Cell Lysate with Human CD45 peptide (ab17553)
Lysates/proteins at 20 µg per lane.
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution
Predicted band size: 147 kDa
Exposure time: 3 minutes
IHC image of CD45 antibody staining in a section of formalin-fixed paraffin-embedded normal human spleen* performed on a Leica BONDTM system using the standard protocol. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab10558, 1ug/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.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
All lanes : Anti-CD45 antibody (ab10558) at 1 µg/ml
Lane 1 : Jurkat (Human) Whole Cell Lysate
Lane 2 : RAW 264.7 (Mouse leukaemic monocyte macrophage cell line) Whole Cell Lysate
Lane 3 : Spleen (Mouse) Tissue Lysate
Lane 4 : Spleen (Rat) Tissue Lysate
Lysates/proteins at 20 µg per lane.
All lanes : Goat Anti-Rabbit IgG H&L (HRP) at 1/50000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 147 kDa
Observed band size: 190 kDa why is the actual band size different from the predicted?
Additional bands at: 230 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 1 minute
This blot was produced using a 3-8% Tris Acetate gel under the TA buffer system. The gel was run at 150V for 60 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab10558 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.
CD45 contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.
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.
Immunohistochemical analysis of formaldehyde fixed human cephalic sections. Primary antibody ab10558 to CD45 incubated at a concentration of 1/100 for 4°C for 18 hours. Secondary antibody used was a goat anti-rabbit congugated to biotin at a 1/200 dilution. Blocking was done with serum at a 10% concentration for 1 hour at 25°C.
ab10558 は 198 報の論文で使用されています。
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