Recombinant Anti-SIRP alpha antibody [EPR22930-163] (ab260039)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22930-163] to SIRP alpha
- Suitable for: ICC/IF, IHC-P, WB, Flow Cyt
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-SIRP alpha antibody [EPR22930-163]
See all SIRP alpha primary antibodies -
Description
Rabbit monoclonal [EPR22930-163] to SIRP alpha -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, IHC-P, WB, Flow Cytmore details
Unsuitable for: IP -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Human brain, U-937, SK-MEL-28 and SK-MEL-2 lysates. IHC-P: Human cerebrum and spleen tissues. ICC/IF: SK-MEL-28 cells. Flow Cyt: THP-1 cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22930-163 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab260039 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 |
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ICC/IF |
1/250.
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IHC-P |
1/500.
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WB |
1/1000. Predicted molecular weight: 55 kDa.
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Flow Cyt |
1/600.
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Notes |
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ICC/IF
1/250. |
IHC-P
1/500. |
WB
1/1000. Predicted molecular weight: 55 kDa. |
Flow Cyt
1/600. |
Target
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Function
Immunoglobulin-like cell surface receptor for CD47. Acts as docking protein and induces translocation of PTPN6, PTPN11 and other binding partners from the cytosol to the plasma membrane. Supports adhesion of cerebellar neurons, neurite outgrowth and glial cell attachment. May play a key role in intracellular signaling during synaptogenesis and in synaptic function (By similarity). Involved in the negative regulation of receptor tyrosine kinase-coupled cellular responses induced by cell adhesion, growth factors or insulin. Mediates negative regulation of phagocytosis, mast cell activation and dendritic cell activation. CD47 binding prevents maturation of immature dendritic cells and inhibits cytokine production by mature dendritic cells. -
Tissue specificity
Ubiquitous. Highly expressed in brain. Detected on myeloid cells, but not T-cells. Detected at lower levels in heart, placenta, lung, testis, ovary, colon, liver, small intestine, prostate, spleen, kidney, skeletal muscle and pancreas. -
Sequence similarities
Contains 2 Ig-like C1-type (immunoglobulin-like) domains.
Contains 1 Ig-like V-type (immunoglobulin-like) domain. -
Post-translational
modificationsN-glycosylated.
Phosphorylated on tyrosine residues in response to stimulation with EGF, growth hormone, insulin and PDGF. Dephosphorylated by PTPN11. -
Cellular localization
Membrane. - Information by UniProt
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Database links
- Entrez Gene: 140885 Human
- Omim: 602461 Human
- SwissProt: P78324 Human
- Unigene: 581021 Human
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Alternative names
- Signal regulatory protein alpha type 1 antibody
- Bit antibody
- Brain Ig like molecule with tyrosine based activation motifs antibody
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of formaldehyde-fixed human lung tissue staining with ab260039 at 1/250 dilution. Sections were cut at 2 µm from lung tissues and mounted on poly-L-lysin coated slides. Samples were incubated with the primary antibody with BSA 5% in PBS for 16 hours at 25°C. Then, the slides were washed in PBS pH 7.4, incubated with secondary antibody Alexa Fluor™ 568 Goat anti-rabbit IgG(H+L) at 1/250 dilution for 30 min at room temperature. Blocking was done using 5% BSA for 20 minutes at 25°C. The second step was performed using primary antibody to CD68 for 1h at room temperature. Then, the slides were washed in PBS pH 7.4, incubated with the appropriate anti mouse secondary antibody 1/250 dilution for 30 min and the DNA was counterstained with Hoechst 33342 at 1µg/ml for 30 min at room temperature. After final washes with PBS, coverslips were mounted on the top of the samples using homemade Mowiol mounting medium.
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Immunohistochemical analysis of paraffin-embedded human spleen tissue labeling SIRP alpha with ab260039 at 1/500 followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on the human spleen is observed. The section was incubated with ab260039 for 15 mins at room temperature. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin.
Secondary antibody only control: Ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.
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All lanes : Anti-SIRP alpha antibody [EPR22930-163] (ab260039) at 1/1000 dilution
Lane 1 : Human brain lysate
Lane 2 : U-937 (human histiocytic lymphoma monocyte) whole cell lysate
Lane 3 : SK-MEL-28 (human malignant melanoma) whole cell lysate
Lane 4 : SK-MEL-2 (human skin malignant melanoma) whole cell lysate
Lane 5 : Jurkat (human T-cell leukemia lymphocyte) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 55 kDa
Observed band size: 85 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST.
There are many forms of SIRP alpha because of different glycosylation and phosphotyrosine modification.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 9712903).
Negative control: Jurkat (PMID: 17982459).
Exposure time: 3 minutes
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Flow cytometric analysis of Jurkat (Human T cell leukemia T lymphocyte, Left) / THP-1 (Human monocytic leukemia monocyte, Right) cells labeling SIRP alpha with ab260039 at 1\600 compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 was used as the secondary antibody.
Negative control: Jurkat.
Gated on viable cells.
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SK-MEL-28 (human malignant melanoma melanoma) cells labeling SIRP alpha with ab260039 at 1/250 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing membranous staining in SK-MEL-28 cells Negative control: Jurkat (PMID: 17982459) is observed. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).
Secondary antibody only control: ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.
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Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling SIRP alpha with ab260039 at 1/500 followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on the human cerebrum (PMID: 9062191) is observed. The section was incubated with ab260039 for 15 mins at room temperature. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin.
Secondary antibody only control: Ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.
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
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SDS download
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Datasheet download
Certificate of Compliance
References (2)
ab260039 has been referenced in 2 publications.
- Yang H et al. SIRPα and PD1 expression on tumor-associated macrophage predict prognosis of intrahepatic cholangiocarcinoma. J Transl Med 20:140 (2022). PubMed: 35317832
- Xu L et al. Identification of macrophage correlated biomarkers to predict the prognosis in patients with intrahepatic cholangiocarcinoma. Front Oncol 12:967982 (2022). PubMed: 36158683