Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-SIRP alpha antibody [EPR16264] (ab191419)

Overview

  • 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, WB, Flow Cytmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human SIRP alpha aa 350-500. The exact sequence is proprietary.
    Database link: P78324

  • Positive control

    • THP1, SW480, C6, PC12, NIH 3T3 and Human fetal brain lysates; THP1 cells. ICC/IF: C6 cells
  • General notes

     

     

    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 rabbit monoclonal antibody.

Properties

Applications

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.
WB 1/1000 - 1/2000. Detects a band of approximately 75-110 kDa (predicted molecular weight: 55 kDa).
Flow Cyt 1/50.

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

Target

  • 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
    modifications

    N-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
  • Database links

  • Alternative names

    • Signal regulatory protein alpha type 1 antibody
    • Bit antibody
    • Brain Ig like molecule with tyrosine based activation motifs antibody
    • Brain Ig-like molecule with tyrosine-based activation motifs antibody
    • Brain immunoglobulin like molecule with tyrosine based activation motifs antibody
    • CD172 antigen like family member A antibody
    • CD172 antigen-like family member A antibody
    • CD172a antibody
    • CD172a antigen antibody
    • Inhibitory receptor SHPS-1 antibody
    • Macrophage fusion receptor antibody
    • MFR antibody
    • MYD 1 antibody
    • Myd 1 antigen antibody
    • MyD-1 antigen antibody
    • p84 antibody
    • Protein tyrosine phosphatase non receptor type substrate 1 antibody
    • PTPNS1 antibody
    • SHP substrate 1 antibody
    • SHPS-1 antibody
    • SHPS1 antibody
    • SHPS1_HUMAN antibody
    • Signal regulatory protein alpha 2 antibody
    • Signal regulatory protein alpha 3 antibody
    • Signal regulatory protein alpha antibody
    • Signal regulatory protein alpha type 2 antibody
    • Signal-regulatory protein alpha-1 antibody
    • Signal-regulatory protein alpha-2 antibody
    • Signal-regulatory protein alpha-3 antibody
    • SIRP antibody
    • Sirp-alpha-1 antibody
    • Sirp-alpha-2 antibody
    • Sirp-alpha-3 antibody
    • SIRPA antibody
    • SIRPalpha antibody
    • SIRPalpha1 antibody
    • SIRPalpha2 antibody
    • SIRPalpha3 antibody
    • Tyrosine phosphatase SHP substrate 1 antibody
    • Tyrosine protein phosphatase non receptor type substrate 1 antibody
    • Tyrosine-protein phosphatase non-receptor type substrate 1 antibody
    see all

Images

  • 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 kDa



    Lanes 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.

  • Immunocytochemistry/ Immunofluorescence analysis of C6 (rat glial tumor glial cell) labeling SIRP alpha with ab191419 at 1/500. ab150077 AlexaFluor®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 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

  • 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

  • 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

References

ab191419 has not yet been referenced specifically in any publications.

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