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Immunohistochemistry (IHC) (Anti-CD45 antibody IHC of human thymus. Immunohistochemistry of formalin-fixed, paraffin-embedded tissue after heat-induced antigen retrieval. Antibody concentration 10 ug/ml.)

Mouse anti-Human CD45 Monoclonal Antibody | anti-CD45 antibody

Mouse Monoclonal [clone 3G4] (IgG2a) to Human CD45

Gene Names
PTPRC; LCA; LY5; B220; CD45; L-CA; T200; CD45R; GP180
Reactivity
Human
Applications
Immunohistochemistry, Western Blot, ELISA
Purity
Protein G Purified
Synonyms
CD45; Monoclonal Antibody; Mouse Monoclonal [clone 3G4] (IgG2a) to Human CD45; Anti-CD45 Antibody (aa1029-1249; clone 3G4) IHC-plus; PTPRC; B220; CD45R; gp180; Leukocyte common antigen; L-CA; Leucocyte common antigen; LY5; T200; T200 glycoprotein; T200 leukocyte common antigen; CD45 antigen; LCA; Human CD45; anti-CD45 antibody
Ordering
For Research Use Only!
Host
Mouse
Reactivity
Human
Clonality
Monoclonal
Isotype
IgG2a
Clone Number
3G4
Purity/Purification
Protein G Purified
Form/Format
PBS, pH 7.4, 0.1% sodium azide
Concentration
1 mg/ml (varies by lot)
Sequence Length
1304
Applicable Applications for anti-CD45 antibody
Immunohistochemistry (IHC - Paraffin), Western Blot (WB), ELISA (EIA)
Application Notes
IHC-P (10 ug/ml)
WB (1:1000 - 1:3000)
Optimal dilutions to be determined by the researcher.

Usage: Immunohistochemistry: MBS245401 was validated for use in immunohistochemistry on a panel of 21 formalin-fixed, paraffin-embedded (FFPE) human tissues after heat induced antigen retrieval in pH 6.0 citrate buffer. After incubation with the primary antibody, slides were incubated with biotinylated secondary antibody, followed by alkaline phosphatase-streptavidin and chromogen. The stained slides were evaluated by a pathologist to confirm staining specificity. The optimal working concentration for MBS245401 was determined to be 10 ug/ml.
Immunogen Species
CD45/LCA antibody was raised against Human
Family/Subfamily
Protein Phosphatase/R1/R6
Antigen Type
Recombinant protein
Immunogen
CD45 antibody was raised against recombinant human CD45 (1029-1249aa) purified from E Coli.
Epitope
aa1029-1249
Disclaimer
Due to the highly specific nature of antibodies and antigens, we cannot predict or be held responsible with respect to how this antibody will behave in your systems. Researchers using this antibody should conduct optimization studies to achieve the most optimal result possible for their intended application.
Recommended Immunohistochemistry Protocol
The following protocol is a recommendation only, and MyBioSource, Inc. makes no guarantee of the results:

Tissue Preparation:
Formalin fixation and embedding in paraffin wax.

Tissue Sectioning:
Make 4-um sections and place on pre-cleaned and charged microscope slides. Heat in a tissue-dryingoven for 45 minutes at 60°C.

Deparaffinization:
Wash dry slides in 3 changes of xylene - 5 minutes each @ RT

Rehydration:
Wash slides in 3 changes of 100% alcohol - 3 minutes each @ RT
Wash slides in 2 changes of 95% alcohol - 3 minutes each @ RT
Wash slides in 1 change of 80% alcohol - 3 minutes @ RT
Rinse slides in gentle running distilled water - 5 minutes @ RT

Antigen retrieval:
Steam slides in 0.01 M sodium citrate buffer, pH 6.0 at 99-100°C - 20 minutes
Remove from heat and let stand at room temperature in buffer - 20 minutes
Rinse in 1X TBS with Tween (TBST) -1 minute @ RT

Immunostaining:
(Do not allow tissues to dry at any time during the staining procedure)
Apply a universal protein block - 20 minutes @ RT
Drain protein block from slides, apply diluted primary antibody - 45 minutes @ RT
Rinse slides in 1 X TBST - 1 minute @ RT
Apply a biotinylated secondary antibody appropriate for the primary antibody - 30 minutes @ RT
Rinse slides in 1X TBST -1 minute @ RT
Apply alkaline phosphatase streptavidin - 30 minutes @ RT
Rinse slides in 1X TBST -1 minute @ RT
Apply alkaline phosphatase chromogen substrate - 30 minutes @ RT
Wash slides in distilled water - 1 minute @ RT

Dehydrate:
(This method should only be used if the chromogen substrate is alcohol insoluble (e.g. Vector Red, DAB)
Wash slides in 2 changes of 80% alcohol - 1 minute each @ RT
Wash slides in 2 changes of 95% alcohol - 1 minute each @ RT
Wash slides in 3 changes of 100% alcohol - 1 minute each @ RT
Wash slides in 3 changes of xylene - 1 minute each @ RT
Apply coverslip
Preparation and Storage
Long term: -20 degree C; Short term: +4 degree C; Avoid freeze-thaw cycles.

Immunohistochemistry (IHC)

(Anti-CD45 antibody IHC of human thymus. Immunohistochemistry of formalin-fixed, paraffin-embedded tissue after heat-induced antigen retrieval. Antibody concentration 10 ug/ml.)

Immunohistochemistry (IHC) (Anti-CD45 antibody IHC of human thymus. Immunohistochemistry of formalin-fixed, paraffin-embedded tissue after heat-induced antigen retrieval. Antibody concentration 10 ug/ml.)

Immunohistochemistry (IHC)

(Anti-CD45 antibody IHC of human tonsil. Immunohistochemistry of formalin-fixed, paraffin-embedded tissue after heat-induced antigen retrieval. Antibody concentration 10 ug/ml.)

Immunohistochemistry (IHC) (Anti-CD45 antibody IHC of human tonsil. Immunohistochemistry of formalin-fixed, paraffin-embedded tissue after heat-induced antigen retrieval. Antibody concentration 10 ug/ml.)

Western Blot (WB)

(Cell lysates of Jurkat(20 ug) were resolved by SDS-PAGE, transferred to NC membrane and probed with anti-human CD45 (1:2000). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and an ECL detection system.)

Western Blot (WB) (Cell lysates of Jurkat(20 ug) were resolved by SDS-PAGE, transferred to NC membrane and probed with anti-human CD45 (1:2000). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and an ECL detection system.)
Related Product Information for anti-CD45 antibody
CD45 (PTPRC) is a family of high-molecular weight glycoproteins uniquely expressed on the surface of all leukocytes and their hematopoietic progenitors. The CD45 family consists of multiple members that are all products of a single gene. Structurally, CD45 is composed of one fibronectin extracellular domain, one transmembrane domain, and two cytoplasmic tyrosine phosphatase domains. It is essential for B- and T-cell activation.
Product Categories/Family for anti-CD45 antibody

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
UniProt Accession #
Molecular Weight
130,898 Da
NCBI Official Full Name
receptor-type tyrosine-protein phosphatase C isoform 1
NCBI Official Synonym Full Names
protein tyrosine phosphatase, receptor type, C
NCBI Official Symbol
PTPRC
NCBI Official Synonym Symbols
LCA; LY5; B220; CD45; L-CA; T200; CD45R; GP180
NCBI Protein Information
receptor-type tyrosine-protein phosphatase C; CD45 antigen; T200 glycoprotein; T200 leukocyte common antigen; protein tyrosine phosphatase, receptor type, c polypeptide
UniProt Protein Name
Receptor-type tyrosine-protein phosphatase C
UniProt Gene Name
PTPRC
UniProt Synonym Gene Names
CD45; L-CA
UniProt Entry Name
PTPRC_HUMAN

NCBI Description

The protein encoded by this gene is a member of the protein tyrosine phosphatase (PTP) family. PTPs are known to be signaling molecules that regulate a variety of cellular processes including cell growth, differentiation, mitosis, and oncogenic transformation. This PTP contains an extracellular domain, a single transmembrane segment and two tandem intracytoplasmic catalytic domains, and thus is classified as a receptor type PTP. This PTP has been shown to be an essential regulator of T- and B-cell antigen receptor signaling. It functions through either direct interaction with components of the antigen receptor complexes, or by activating various Src family kinases required for the antigen receptor signaling. This PTP also suppresses JAK kinases, and thus functions as a regulator of cytokine receptor signaling. Alternatively spliced transcripts variants of this gene, which encode distinct isoforms, have been reported. [provided by RefSeq, Jun 2012]

Uniprot Description

CD45: a receptor type protein tyrosine phosphatase protein. Contains an extracellular domain, a single transmembrane segment and two tandem intracytoplasmic catalytic domains. The first PTPAse domain has enzymatic activity, while the second one seems to affect the substrate specificity of the first one. Specifically expressed in hematopoietic cells. An essential regulator of T- and B-cell antigen receptor signaling. Functions through either direct interaction with components of the antigen receptor complexes, or by activating various Src family kinases required for the antigen receptor signaling. Suppresses JAK kinases, and thus functions as a regulator of cytokine receptor signaling. Four alternatively spliced isoforms have been described.

Protein type: Membrane protein, integral; Motility/polarity/chemotaxis; EC 3.1.3.48; Receptor protein phosphatase, tyrosine

Chromosomal Location of Human Ortholog: 1q31-q32

Cellular Component: focal adhesion; membrane; integral to plasma membrane; plasma membrane; lipid raft; external side of plasma membrane

Molecular Function: protein binding; transmembrane receptor protein tyrosine phosphatase activity; protein tyrosine phosphatase activity; protein kinase binding

Biological Process: B cell proliferation; axon guidance; substrate-bound cell migration, cell release from substrate; regulation of cell cycle; protein amino acid dephosphorylation; positive regulation of antigen receptor-mediated signaling pathway; T cell receptor signaling pathway; bone marrow development; B cell receptor signaling pathway; stem cell development; cell surface receptor linked signal transduction; dephosphorylation; release of sequestered calcium ion into cytosol; positive regulation of protein kinase activity; negative regulation of T cell mediated cytotoxicity; positive regulation of B cell proliferation; negative regulation of protein kinase activity; hemopoietic progenitor cell differentiation; positive regulation of T cell proliferation; negative regulation of cytokine and chemokine mediated signaling pathway; immunoglobulin biosynthetic process; defense response to virus; T cell differentiation

Disease: Hepatitis C Virus, Susceptibility To; Severe Combined Immunodeficiency, Autosomal Recessive, T Cell-negative, B Cell-positive, Nk Cell-positive

Research Articles on CD45

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Product Notes

The CD45 ptprc (Catalog #AAA245401) is an Antibody produced from Mouse and is intended for research purposes only. The product is available for immediate purchase. The Mouse Monoclonal [clone 3G4] (IgG2a) to Human CD45 reacts with Human and may cross-react with other species as described in the data sheet. AAA Biotech's CD45 can be used in a range of immunoassay formats including, but not limited to, Immunohistochemistry (IHC - Paraffin), Western Blot (WB), ELISA (EIA). IHC-P (10 ug/ml) WB (1:1000 - 1:3000) Optimal dilutions to be determined by the researcher. Usage: Immunohistochemistry: MBS245401 was validated for use in immunohistochemistry on a panel of 21 formalin-fixed, paraffin-embedded (FFPE) human tissues after heat induced antigen retrieval in pH 6.0 citrate buffer. After incubation with the primary antibody, slides were incubated with biotinylated secondary antibody, followed by alkaline phosphatase-streptavidin and chromogen. The stained slides were evaluated by a pathologist to confirm staining specificity. The optimal working concentration for MBS245401 was determined to be 10 ug/ml. Researchers should empirically determine the suitability of the CD45 ptprc for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "CD45, Monoclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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