Recombinant Anti-SIRP alpha antibody [EPR16264] (ab191419)
![Western blot - Anti-SIRP alpha antibody [EPR16264] (ab191419)](https://www.abcam.com/ps/products/191/ab191419/Images/ab191419-477279-anti-sirp-alpha-antibody-epr16264-western-blot-wildtype-raw-2647-sirpa-knockout-raw-2647-thp1-mcf7.jpg "Western blot - Anti-SIRP alpha antibody [EPR16264] (ab191419)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR16264] to SIRP alpha
- Suitable for: ICC/IF, Flow Cyt (Intra), WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-SIRP alpha antibody [EPR16264]
See all SIRP alpha primary antibodies -
Description
Rabbit monoclonal [EPR16264] to SIRP alpha -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, Flow Cyt (Intra), WBmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: THP1, SW480, C6, PC12, NIH 3T3 and Human fetal brain lysates. Flow Cyt (intra): THP1 cells. ICC/IF: C6 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: 40% Glycerol (glycerin, glycerine), 0.05% BSA, 59% PBS -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR16264 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab191419 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 |
|---|---|---|
| ICC/IF |
1/500.
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| Flow Cyt (Intra) |
1/50.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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| WB |
1/1000 - 1/2000. Detects a band of approximately 75-110 kDa (predicted molecular weight: 55 kDa).
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| Notes |
|---|
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ICC/IF
1/500. |
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Flow Cyt (Intra)
1/50. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB
1/1000 - 1/2000. Detects a band of approximately 75-110 kDa (predicted molecular weight: 55 kDa). |
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
- Entrez Gene: 19261 Mouse
- Entrez Gene: 25528 Rat
- Omim: 602461 Human
- SwissProt: P78324 Human
- SwissProt: P97797 Mouse
- SwissProt: P97710 Rat
- Unigene: 581021 Human
see all -
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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Western blot - Anti-SIRP alpha antibody [EPR16264] (ab191419)All lanes : Anti-SIRP alpha antibody [EPR16264] (ab191419) at 1/1000 dilution
Lane 1 : Wild-type RAW 264.7 cell lysate
Lane 2 : SIRPA knockout RAW 264.7 cell lysate
Lane 3 : THP-1 cell lysate
Lane 4 : MCF7 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 55 kDa
Observed band size: 100-140 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-SIRP alpha antibody [EPR16264] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab191419 was shown to bind specifically to SIRP alpha. A band was observed at 100-140 kDa (mouse SIRPA, isoform 1) & 40-50 kDa (mouse SIRPA, isoform 2), in wild-type RAW 264.7 cell lysates (band observed at 70-100 kDa in THP-1 is Human SIRPA) with no signal observed at this size in SIRPA knockout cell line ab281618 (knockout cell lysate ab282969). To generate this image, wild-type and SIRPA knockout RAW 264.7 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
Glycosylation level (~65-120 kDa) of SIRPα is different in various tissues (PMID: 18051954).
Observed band: 100-140 kDa (mouse SIRPA, isoform 1) & 40-50 kDa (mouse SIRPA, isoform 2).
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![Western blot - Anti-SIRP alpha antibody [EPR16264] (ab191419)](https://www.abcam.com/ps/products/191/ab191419/Images/ab191419-312620-anti-sirp-alpha-antibody-epr16264-western-blot.jpg)
Western blot - Anti-SIRP alpha antibody [EPR16264] (ab191419)All lanes : Anti-SIRP alpha antibody [EPR16264] (ab191419) at 1/1000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : SIRPA knockout HAP1 whole cell lysate
Lane 3 : THP1 whole cell lysate
Lane 4 : Jurkat whole cell lysate (negative control)
Lysates/proteins at 20 µg per lane.
Predicted band size: 55 kDa
Observed band size: 55 kDaLanes 1 - 4: Merged signal (red and green). Green - ab191419 observed at 55 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab191419 was shown to recognize SIRP alpha in wild-type HAP1 cells as signal was lost at the expected MW in SIRPA knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and SIRPA knockout samples were subjected to SDS-PAGE. Ab191419 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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![Immunocytochemistry/ Immunofluorescence - Anti-SIRP alpha antibody [EPR16264] (ab191419)](https://www.abcam.com/ps/products/191/ab191419/Images/ab191419-343547-antiSIRPalphaantibodyEPR16264-immunocytochemistryimmunofluorescence-c6-rat.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-SIRP alpha antibody [EPR16264] (ab191419)Immunocytochemistry/ Immunofluorescence analysis of C6 (rat glial tumor glial cell) labeling SIRP alpha with ab191419 at 1/500. ab150077 Alexa Fluor® 488 Goat anti-Rabbit at 1/1000 was used as the secondary antibody. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. DAPI was used to stain nuclei blue.Confocal image showing membranous staining on C6 cell line. -
![Flow Cytometry (Intracellular) - Anti-SIRP alpha antibody [EPR16264] (ab191419)](https://www.abcam.com/ps/products/191/ab191419/Images/ab191419-230818-anti-sirp-alpha-antibody-epr16264-flow-cytometry.jpg)
Flow Cytometry (Intracellular) - Anti-SIRP alpha antibody [EPR16264] (ab191419)Intracellular flow cytometric analysis of THP1 cells (2% paraformaldehyde-fixed) labeling SIRP alpha with ab191419 at 1/50 dilution (red) or a Rabbit monoclonal IgG (negative) (green), followed by Goat anti rabbit IgG (FITC) secondary at 1/150 dilution
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![Western blot - Anti-SIRP alpha antibody [EPR16264] (ab191419)](https://www.abcam.com/ps/products/191/ab191419/Images/ab191419-230820-anti-sirp-alpha-antibody-epr16264-western-blot.jpg)
Western blot - Anti-SIRP alpha antibody [EPR16264] (ab191419)All lanes : Anti-SIRP alpha antibody [EPR16264] (ab191419) at 1/5000 dilution
Lane 1 : THP1 cell lysate
Lane 2 : SW480 cell lysate
Lane 3 : Human fetal brain lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 55 kDa -
![Western blot - Anti-SIRP alpha antibody [EPR16264] (ab191419)](https://www.abcam.com/ps/products/191/ab191419/Images/ab191419-230819-anti-sirp-alpha-antibody-epr16264-western-blot.jpg)
Western blot - Anti-SIRP alpha antibody [EPR16264] (ab191419)All lanes : Anti-SIRP alpha antibody [EPR16264] (ab191419) at 1/1000 dilution
Lane 1 : C6 cell lysate
Lane 2 : PC12 cell lysate
Lane 3 : NIH 3T3 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 55 kDa -
![Anti-SIRP alpha antibody [EPR16264] (ab191419)](https://www.abcam.com/ps/products/191/ab191419/Images/ab191419-6-benefits-of-recombinant-antibodies.png)
Anti-SIRP alpha antibody [EPR16264] (ab191419)
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (3)
ab191419 has been referenced in 3 publications.
- Wu Y et al. The disbalance of LRP1 and SIRPα by psychological stress dampens the clearance of tumor cells by macrophages. Acta Pharm Sin B 12:197-209 (2022). PubMed: 35127380
- Wang X et al. MicroRNA-382 Promotes M2-Like Macrophage via the SIRP-α/STAT3 Signaling Pathway in Aristolochic Acid-Induced Renal Fibrosis. Front Immunol 13:864984 (2022). PubMed: 35585990
- Faralli JA et al. Genomic/proteomic analyses of dexamethasone-treated human trabecular meshwork cells reveal a role for GULP1 and ABR in phagocytosis. Mol Vis 25:237-254 (2019). PubMed: 31516309