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Rabbit Recombinant Monoclonal SIRP alpha antibody. Carrier free. Suitable for WB, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse, Rat samples.

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Images

Western blot - Anti-SIRP alpha antibody [EPR16264] - Low endotoxin, Azide free (AB209892), expandable thumbnail
  • Western blot - Anti-SIRP alpha antibody [EPR16264] - Low endotoxin, Azide free (AB209892), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-SIRP alpha antibody [EPR16264] - Low endotoxin, Azide free (AB209892), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-SIRP alpha antibody [EPR16264] - Low endotoxin, Azide free (AB209892), expandable thumbnail

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Constituents: PBS

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
WBICC/IFFlow Cyt (Intra)
Human
Tested
Expected
Tested
Mouse
Tested
Expected
Expected
Rat
Tested
Tested
Expected

Tested
Tested

Species

Human, Mouse, Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Rat

Dilution info

-

Notes

-

Expected
Expected

Species

Human, Mouse

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Associated Products

Select an associated product type

12 products for Alternative Product

4 products for Alternative Version

Target data

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.

Alternative names

Tyrosine-protein phosphatase non-receptor type substrate 1, SHP substrate 1, SHPS-1, Brain Ig-like molecule with tyrosine-based activation motifs, CD172 antigen-like family member A, Inhibitory receptor SHPS-1, Macrophage fusion receptor, MyD-1 antigen, Signal-regulatory protein alpha-1, Signal-regulatory protein alpha-2, Signal-regulatory protein alpha-3, p84, Bit, Sirp-alpha-1, Sirp-alpha-2, Sirp-alpha-3, BIT, MFR, SIRP, SHPS1, PTPNS1, MYD1, SIRPA

Recommended products

Rabbit Recombinant Monoclonal SIRP alpha antibody. Carrier free. Suitable for WB, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse, Rat samples.

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Carrier free

Yes

Clone number

EPR16264

Purification technique

Affinity purification Protein A

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate long-term storage conditions

+4°C

Storage information

Do Not Freeze

Notes

ab209892 is the carrier-free version of Anti-SIRP alpha antibody [EPR16264] ab191419.

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

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 batch production

For more information, read more on recombinant antibodies.

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.

Our Low endotoxin, azide-free formats have low endotoxin level (1 EU/mg, determined by the TAL assay) and are free from azide, to achieve consistent experimental results in functional assays.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

SIRP alpha also known as CD172a is a transmembrane receptor protein with a mass ranging between 70-110 kDa due to glycosylation. This protein extensively expresses on the surface of myeloid cells neurons and a subset of T-cells and is part of the immunoglobulin superfamily. SIRP alpha interacts with its ligand CD47 a widely expressed glycoprotein involved in immune response regulation. Its mechanical action primarily involves signal regulation through the recruitment of SHP-1 and SHP-2 two cytoplasmic tyrosine phosphatases.

Biological function summary

SIRP alpha functions significantly in the regulation of phagocytosis acting as a "don't eat me" signal to macrophages upon binding with CD47. It does not act alone; rather it is part of a complex that recruits SHP-1 and SHP-2 leading to inhibition of dephosphorylation activities essential for engulfment processes. This regulatory mechanism is important for maintaining cellular homeostasis ensuring that healthy cells are not mistakenly destroyed by the immune system.

Pathways

SIRP alpha plays an important role in the innate immune pathways involving the regulation of phagocytosis and cell-cell adhesion. Particularly it fits into the immune checkpoint pathways where it interacts closely with proteins like CD47 and plays a role in the interaction between the immune system and cancer cells. Through these pathways SIRP alpha helps maintain balance in the immune response allowing for the recognition of self versus non-self therefore preventing autoimmunity while facilitating the clearance of pathogens.

Associated diseases and disorders

SIRP alpha is implicated in cancer and autoimmune diseases. In cancer its interaction with CD47 allows tumor cells to escape phagocytosis promoting tumor survival and growth. In autoimmune disorders dysregulated SIRP alpha expression or signaling could miscommunicate immune signals leading to the destruction of healthy tissue. Understanding the link between SIRP alpha and these conditions can reveal potential targets for therapeutic development especially using inhibitors or modulators targeting the SIRP alpha-CD47 interaction.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

4 product images

  • Western blot - Anti-SIRP alpha antibody [EPR16264] - Low endotoxin, Azide free (ab209892), expandable thumbnail

    Western blot - Anti-SIRP alpha antibody [EPR16264] - Low endotoxin, Azide free (ab209892)

    False colour image of Western blot: Anti-SIRP alpha antibody [EPR16264] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-SIRP alpha antibody [EPR16264] 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 Mouse SIRPA knockout RAW 264.7 cell line ab281618 (knockout cell lysate Mouse SIRPA knockout RAW 264.7 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed 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)

    All lanes: Western blot - Anti-SIRP alpha antibody [EPR16264] (Anti-SIRP alpha antibody [EPR16264] ab191419) at 1/1000 dilution

    Lane 1: Wild-type RAW 264.7 cell lysate at 20 µg

    Lane 2: SIRPA knockout RAW 264.7 cell lysate at 20 µg

    Lane 3: THP-1 cell lysate at 20 µg

    Lane 4: MCF7 cell lysate at 20 µg

    Performed under reducing conditions.

    Predicted band size: 55 kDa

    Observed band size: 100-140 kDa

  • Western blot - Anti-SIRP alpha antibody [EPR16264] - Low endotoxin, Azide free (ab209892), expandable thumbnail

    Western blot - Anti-SIRP alpha antibody [EPR16264] - Low endotoxin, Azide free (ab209892)

    Lanes 1 - 4: Merged signal (red and green). Green - Anti-SIRP alpha antibody [EPR16264] ab191419 observed at 55 kDa. Red - loading control, Anti-GAPDH antibody [mAbcam 9484] - Loading Control ab9484, observed at 37 kDa.

    Anti-SIRP alpha antibody [EPR16264] 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. Anti-SIRP alpha antibody [EPR16264] ab191419 and Anti-GAPDH antibody [mAbcam 9484] - Loading Control 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 Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-SIRP alpha antibody [EPR16264] ab191419).

    All lanes: Western blot - Anti-SIRP alpha antibody [EPR16264] (Anti-SIRP alpha antibody [EPR16264] ab191419) at 1/1000 dilution

    Lane 1: Wild-type HAP1 whole cell lysate at 20 µg

    Lane 2: SIRPA knockout HAP1 whole cell lysate at 20 µg

    Lane 3: THP1 whole cell lysate at 20 µg

    Lane 4: Jurkat whole cell lysate (negative control) at 20 µg

    Predicted band size: 55 kDa

    Observed band size: 55 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-SIRP alpha antibody [EPR16264] - Low endotoxin, Azide free (ab209892), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-SIRP alpha antibody [EPR16264] - Low endotoxin, Azide free (ab209892)

    Immunocytochemistry/ Immunofluorescence analysis of C6 (rat glial tumor glial cell) labeling SIRP alpha with Anti-SIRP alpha antibody [EPR16264] ab191419 at 1/500. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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 (Anti-SIRP alpha antibody [EPR16264] ab191419).

  • Flow Cytometry (Intracellular) - Anti-SIRP alpha antibody [EPR16264] - Low endotoxin, Azide free (ab209892), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-SIRP alpha antibody [EPR16264] - Low endotoxin, Azide free (ab209892)

    Intracellular flow cytometric analysis of THP1 cells (2% paraformaldehyde-fixed) labeling SIRP alpha with Anti-SIRP alpha antibody [EPR16264] 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 (Anti-SIRP alpha antibody [EPR16264] ab191419).

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Product protocols

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