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RecombinantRabMAb

Recombinant Anti-SIRP alpha antibody [EPR24187-17] (ab259357)

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Western blot - Anti-SIRP alpha antibody [EPR24187-17] (ab259357)
  • Western blot - Anti-SIRP alpha antibody [EPR24187-17] (ab259357)
  • Flow Cytometry (Intracellular) - Anti-SIRP alpha antibody [EPR24187-17] (ab259357)
  • Immunocytochemistry - Anti-SIRP alpha antibody [EPR24187-17] (ab259357)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRP alpha antibody [EPR24187-17] (ab259357)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRP alpha antibody [EPR24187-17] (ab259357)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRP alpha antibody [EPR24187-17] (ab259357)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRP alpha antibody [EPR24187-17] (ab259357)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRP alpha antibody [EPR24187-17] (ab259357)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRP alpha antibody [EPR24187-17] (ab259357)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR24187-17] to SIRP alpha
  • Suitable for: Flow Cyt (Intra), ICC/IF, WB, IHC-P
  • Reacts with: Mouse, Rat

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-SIRP alpha antibody [EPR24187-17]
    See all SIRP alpha primary antibodies

  • Description

    Rabbit monoclonal [EPR24187-17] to SIRP alpha

  • Host species

    Rabbit

  • Specificity

    ICC and Flow Cyt applications do not react with Rat species.

  • Tested applications

    Suitable for: Flow Cyt (Intra), ICC/IF, WB, IHC-Pmore details
    Unsuitable for: IP

  • Species reactivity

    Reacts with: Mouse, Rat

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Mouse cerebellum, Rat hippocampus and RAW264.7 lysates. IHC-P: Mouse cerebrum, Mouse spleen, Mouse lung carcinoma, Rat cerebrum and Rat spleen tissues. ICC: RAW264.7 cells. Flow cyt (Intra): RAW264.7 cells.

  • 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
    For more information see here.

    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

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab259357 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)
1/50.
ICC/IF
1/100.
WB
1/1000. Predicted molecular weight: 55 kDa.
IHC-P
1/1000.
Notes
Flow Cyt (Intra)
1/50.
ICC/IF
1/100.
WB
1/1000. Predicted molecular weight: 55 kDa.
IHC-P
1/1000.
Application notes
Is unsuitable for IP.

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

    • Western blot - Anti-SIRP alpha antibody [EPR24187-17] (ab259357)
      Western blot - Anti-SIRP alpha antibody [EPR24187-17] (ab259357)
      All lanes : Anti-SIRP alpha antibody [EPR24187-17] (ab259357) at 1/1000 dilution

      Lane 1 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
      Lane 2 : A20 (mouse reticum sarcoma b lymphocyte) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/5000 dilution

      Predicted band size: 55 kDa
      Observed band size: 120 kDa why is the actual band size different from the predicted?



      Blocking and diluting buffer and concentration: 5% NFDM/TBST. The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 24143245). 

      Negative control: A20 (PMID:9712903).

      Exposure time: 70 seconds

       

    • Western blot - Anti-SIRP alpha antibody [EPR24187-17] (ab259357)
      Western blot - Anti-SIRP alpha antibody [EPR24187-17] (ab259357)
      All lanes : Anti-SIRP alpha antibody [EPR24187-17] (ab259357) at 1/1000 dilution

      Lane 1 : Mouse cerebellum tissue lysate
      Lane 2 : Rat hippocampus tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 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. The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID:28545141).

      Exposure time: Lane 1: 125 seconds; Lane 2: 3 minutes

       

    • Flow Cytometry (Intracellular) - Anti-SIRP alpha antibody [EPR24187-17] (ab259357)
      Flow Cytometry (Intracellular) - Anti-SIRP alpha antibody [EPR24187-17] (ab259357)

      Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized A20 (Mouse reticulum sarcoma B lymphocyte, Left) / RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage, Right) cells labelling SIRP alpha with ab259357 at 1/50 dilution (1ug)/ (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

    • Immunocytochemistry - Anti-SIRP alpha antibody [EPR24187-17] (ab259357)
      Immunocytochemistry - Anti-SIRP alpha antibody [EPR24187-17] (ab259357)

      Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 cells labelling SIRP alpha with ab259357 at 1/100 (4.41 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing membranous and cytoplasmic staining in RAW 264.7 cell line. Negative control: A20 (PMID: 9712903 . ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue).

      Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRP alpha antibody [EPR24187-17] (ab259357)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRP alpha antibody [EPR24187-17] (ab259357)

      Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labelling SIRP alpha with ab259357 at 1/1000 (0.441 ug/ml) followed by a ready to use LeicaDS9800 (BOND™, Polymer Refine Detection) was used. Negative control: no staining on mouse skeletal muscle. The section was incubated with ab259357 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (BOND™, Polymer Refine Detection) was used.

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRP alpha antibody [EPR24187-17] (ab259357)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRP alpha antibody [EPR24187-17] (ab259357)

      Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labelling SIRP alpha with ab259357 at 1/1000 (0.441 ug/ml) followed by a ready to use Leica DS9800 (BOND™, Polymer Refine Detection) was used. Positive staining on rat spleen. The section was incubated with ab259357 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use Leica DS9800 (BOND™, Polymer Refine Detection) was used.

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRP alpha antibody [EPR24187-17] (ab259357)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRP alpha antibody [EPR24187-17] (ab259357)

      Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labelling SIRP alpha with ab259357 at 1/1000 (0.441 ug/ml) followed by a ready to use Leica DS9800 (BOND™, Polymer Refine Detection) was used. Positive staining on rat cerebrum. The section was incubated with ab259357 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use Leica DS9800 (BOND™, Polymer Refine Detection) was used.

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRP alpha antibody [EPR24187-17] (ab259357)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRP alpha antibody [EPR24187-17] (ab259357)

      Immunohistochemical analysis of paraffin-embedded Mouse lung carcinoma tissue labelling SIRP alpha with ab259357 at 1/1000 (0.441 ug/ml) followed by a ready to use Leica DS9800 (BOND™, Polymer Refine Detection) was used. Positive staining on mouse lung carcinoma. The section was incubated with ab259357 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use Leica DS9800 (BOND™, Polymer Refine Detection) was used.

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRP alpha antibody [EPR24187-17] (ab259357)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRP alpha antibody [EPR24187-17] (ab259357)

      Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labelling SIRP alpha with ab259357 at 1/1000 (0.441 ug/ml) followed by a ready to use Leica DS9800 (BOND™, Polymer Refine Detection) was used. Positive staining on mouse spleen. The section was incubated with ab259357 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use Leica DS9800 (BOND™, Polymer Refine Detection) was used.

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRP alpha antibody [EPR24187-17] (ab259357)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRP alpha antibody [EPR24187-17] (ab259357)

      Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labelling SIRP alpha with ab259357 at 1/1000 (0.441 ug/ml) followed by a ready to use Leica DS9800 (BOND™, Polymer Refine Detection) was used. Positive staining on mouse cerebrum. The section was incubated with ab259357 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (BOND™, Polymer Refine Detection) was used.

      Heat mediated antigen retrieval 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.

    Click here to view the general protocols

    References (0)

    Publishing research using ab259357? Please let us know so that we can cite the reference in this datasheet.

    ab259357 has not yet been referenced specifically in any publications.

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