Recombinant Anti-SIRP alpha antibody [EPR16264] - BSA and Azide free (ab271957)
![Western blot - Anti-SIRP alpha antibody [EPR16264] - BSA and Azide free (ab271957)](https://www.abcam.com/ps/products/271/ab271957/Images/ab271957-477310-anti-sirp-alpha-antibody-epr16264-bsa-and-azide-free-western-blot-wildtype-raw-2647-sirpa-knockout-raw-2647-thp1-mcf7.jpg "Western blot - Anti-SIRP alpha antibody [EPR16264] - BSA and Azide free (ab271957)")
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 - BSA and Azide free
- Suitable for: Flow Cyt (Intra), WB, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-SIRP alpha antibody [EPR16264] - BSA and Azide free
See all SIRP alpha primary antibodies -
Description
Rabbit monoclonal [EPR16264] to SIRP alpha - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, ICC/IFmore 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
- Flow Cyt (intra): THP1 cells. ICC/IF: C6 cells
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General notes
ab271957 is the carrier-free version of ab191419.
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.
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. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
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
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab271957 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 |
|---|---|---|
| Flow Cyt (Intra) |
Use at an assay dependent concentration.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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| WB |
Use at an assay dependent concentration. Predicted molecular weight: 55 kDa.
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| ICC/IF |
Use at an assay dependent concentration.
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| Notes |
|---|
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Flow Cyt (Intra)
Use at an assay dependent concentration. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
|
WB
Use at an assay dependent concentration. Predicted molecular weight: 55 kDa. |
|
ICC/IF
Use at an assay dependent concentration. |
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] - BSA and Azide free (ab271957)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 kDaFalse 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).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab191419).
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![Immunocytochemistry/ Immunofluorescence - Anti-SIRP alpha antibody [EPR16264] - BSA and Azide free (ab271957)](https://www.abcam.com/ps/products/271/ab271957/Images/ab271957-2-ab191419343547antiSIRPalphaantibodyEPR16264immunocytochemistryimmunofluorescencec6rat.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-SIRP alpha antibody [EPR16264] - BSA and Azide free (ab271957)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.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab191419). -
![Flow Cytometry (Intracellular) - Anti-SIRP alpha antibody [EPR16264] - BSA and Azide free (ab271957)](https://www.abcam.com/ps/products/271/ab271957/Images/ab271957-1-ab191419230818antisirpalphaantibodyepr16264flowcytometry.jpg)
Flow Cytometry (Intracellular) - Anti-SIRP alpha antibody [EPR16264] - BSA and Azide free (ab271957)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
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab191419).
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![Anti-SIRP alpha antibody [EPR16264] - BSA and Azide free (ab271957)](https://www.abcam.com/ps/products/271/ab271957/Images/ab271957-3-benefits-of-recombinant-antibodies.png)
Anti-SIRP alpha antibody [EPR16264] - BSA and Azide free (ab271957)
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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Datasheet download
Certificate of Compliance
References (0)
ab271957 has not yet been referenced specifically in any publications.
