Anti-CD45 antibody [F10-89-4] - BSA and Azide free (ab230296)
Key features and details
- Mouse monoclonal [F10-89-4] to CD45 - BSA and Azide free
- Suitable for: IHC-P, Flow Cyt, ICC/IF
- Reacts with: Human
- Isotype: IgG2a
Related conjugates and formulations
Overview
-
Product name
Anti-CD45 antibody [F10-89-4] - BSA and Azide free
See all CD45 primary antibodies -
Description
Mouse monoclonal [F10-89-4] to CD45 - BSA and Azide free -
Host species
Mouse -
Specificity
Clone F10-89-4 reacts with all forms of CD45 expressed by all haematopoietic cells, except erythrocytes, having a higher level of expression on lymphocytes than on granulocytes.
-
Tested applications
Suitable for: IHC-P, Flow Cyt, ICC/IFmore details -
Species reactivity
Reacts with: Human -
Immunogen
Tissue, cells or virus. This information is proprietary to Abcam and/or its suppliers.
-
Positive control
- IHC-P: Human tonsil (normal) tissue. ICC/IF: Jurkat cells. Flow Cyt: Jurkat cells, whole blood.
-
General notes
ab230296 is the carrier-free version of ab30470.
Clone F10-89-4 reacts with all forms of CD45 expressed by all haematopoietic cells, except erythrocytes, having a higher level of expression on lymphocytes than on granulocytes.
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at +4°C. Do Not Freeze. -
Storage buffer
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
-
Purity
IgG fraction -
Clonality
Monoclonal -
Clone number
F10-89-4 -
Myeloma
NS1 -
Isotype
IgG2a -
Research areas
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab230296 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 |
---|---|---|
IHC-P |
Use a concentration of 1 - 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
|
|
Flow Cyt |
1/10 - 1/50.
ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody. |
|
ICC/IF |
Use a concentration of 5 - 10 µg/ml.
|
Notes |
---|
IHC-P
Use a concentration of 1 - 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
Flow Cyt
1/10 - 1/50. ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody. |
ICC/IF
Use a concentration of 5 - 10 µg/ml. |
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
modificationsHeavily N- and O-glycosylated. -
Cellular localization
Membrane. Membrane raft. Colocalized with DPP4 in membrane rafts. - Information by UniProt
-
Database links
- Entrez Gene: 5788 Human
- Omim: 151460 Human
- SwissProt: P08575 Human
- Unigene: 654514 Human
-
Alternative names
- B220 antibody
- CD 45 antibody
- CD45 antibody
see all
Images
-
ab30470 staining CD45 in Jurkat (Human T cell leukemia cell line from peripheral blood) cells. The cells were fixed with 4% formaldehyde (10 minutes), then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1 hour. The cells were then incubated overnight at +4°C with ab30470 at 5 µg/ml (shown in green) and ab206369, Rabbit monoclonal to beta Tubulin (Alexa Fluor® 594), at 1/250 dilution (shown in red). This was followed by an incubation at room temperature for 1 hour with ab150117, Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed, at 1 µg/ml (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
This product also gave a positive signal under the same testing conditions in Jurkat cells fixed with 80% methanol (5 minutes).
This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab30470).
-
Flow cytometry staining of human whole blood with ab230296 (right) or mouse IgG2aκ; (ab18413) isotype (left). Red blood cells of 200 µL blood were lysed, then cells were incubated for 30 min on ice in 1x PBS containing 10 µg/ml human IgG and 10 % normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab230296) or mouse IgG2aκ; (ab18413) isotype (1x106 in 100 µL at 0.2 µg/ml) for 30 min on ice.
The secondary antibody Goat anti-mouse IgG H&L (Alexa Fluor ® 488, pre-adsorbed) (ab150117) was used at 1/2000 dilution for 30 min on ice.
Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter. Events were gated on alive cells.
-
Overlay histogram showing Jurkat (Human T cell leukemia cell line from peripheral blood) cells stained with ab30470 (red line). The cells were fixed with 4% paraformaldehyde (10 minutes) 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 (ab30470, 1 µg/1 x 106 cells) for 30 minutes at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 minutes at 22ºC. Isotype control antibody (black line) was mouse IgG2a (ab91361 µg/1 x 106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Jurkat cells fixed with 80% methanol (5 minutes) used under the same conditions.
Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab30470).
-
IHC image of CD45 staining in human tonsil 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 (pH 6, epitope retrieval solution 1) for 20 minutes. The section was then incubated with ab30470, 1 µg/ml, for 15 minutes 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab30470).
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
-
Datasheet download
References (0)
ab230296 has not yet been referenced specifically in any publications.