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Rabbit Recombinant Monoclonal SIRP alpha antibody. Suitable for WB, ICC/IF, Flow Cyt (Intra) and reacts with Rat, Mouse, Human samples. Cited in 5 publications.


Images

Western blot - Anti-SIRP alpha antibody [EPR16264] (AB191419), expandable thumbnail
  • Western blot - Anti-SIRP alpha antibody [EPR16264] (AB191419), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-SIRP alpha antibody [EPR16264] (AB191419), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-SIRP alpha antibody [EPR16264] (AB191419), expandable thumbnail
  • Western blot - Anti-SIRP alpha antibody [EPR16264] (AB191419), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

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

Rat

Dilution info

1/1000 - 1/2000

Notes

-

Species

Mouse

Dilution info

1/1000 - 1/2000

Notes

-

Species

Human

Dilution info

1/1000 - 1/2000

Notes

-

Tested
Tested

Species

Rat

Dilution info

1/500

Notes

-

Expected
Expected

Species

Mouse, Human

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Human

Dilution info

1/50

Notes

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

Expected
Expected

Species

Rat, Mouse

Dilution info

Use at an assay dependent concentration.

Notes

-

Associated Products

Select an associated product type

10 products for Alternative Product

1 product 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. Suitable for WB, ICC/IF, Flow Cyt (Intra) and reacts with Rat, Mouse, Human samples. Cited in 5 publications.

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

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

EPR16264

Purification technique

Affinity purification Protein A

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

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 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.

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.

7 product images

  • Western blot - Anti-SIRP alpha antibody [EPR16264] (ab191419), expandable thumbnail

    Western blot - Anti-SIRP alpha antibody [EPR16264] (ab191419)

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

    All lanes: Western blot - 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

  • Western blot - Anti-SIRP alpha antibody [EPR16264] (ab191419), expandable thumbnail

    Western blot - Anti-SIRP alpha antibody [EPR16264] (ab191419)

    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, 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] (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

  • Immunocytochemistry/ Immunofluorescence - Anti-SIRP alpha antibody [EPR16264] (ab191419), expandable thumbnail

    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. 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.

  • Flow Cytometry (Intracellular) - Anti-SIRP alpha antibody [EPR16264] (ab191419), expandable thumbnail

    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

  • Western blot - Anti-SIRP alpha antibody [EPR16264] (ab191419), expandable thumbnail

    Western blot - Anti-SIRP alpha antibody [EPR16264] (ab191419)

    All lanes: Western blot - Anti-SIRP alpha antibody [EPR16264] (ab191419) at 1/5000 dilution

    Lane 1: THP1 cell lysate at 20 µg

    Lane 2: SW480 cell lysate at 20 µg

    Lane 3: Human fetal brain lysate at 20 µg

    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), expandable thumbnail

    Western blot - Anti-SIRP alpha antibody [EPR16264] (ab191419)

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

    Lane 1: C6 cell lysate at 10 µg

    Lane 2: PC12 cell lysate at 10 µg

    Lane 3: NIH 3T3 cell lysate at 10 µg

    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), expandable thumbnail

    Western blot - Anti-SIRP alpha antibody [EPR16264] (ab191419)

    Western blot: Anti-SIRPA antibody [EPR16264] (ab191419) staining at 1/1500 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 magenta. In Western blot, ab191419 was shown to bind specifically to SIRPA. A band was observed at 75-98 kDa in wild-type A549 cell lysates with no signal observed at this size in SIRPA knockout cell line. To generate this image, wild-type and SIRPA knockout A549 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 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

    All lanes: Western blot - Anti-SIRP alpha antibody [EPR16264] (ab191419) at 1/1500 dilution

    Lane 1: Wild-type A549 cell lysate at 20 µg

    Lane 2: SIRPA knockout A549 cell lysate at 20 µg

    Lane 3: THP-1 cell lysate at 20 µg

    Lane 4: SW480 cell lysate at 20 µg

    Secondary

    All lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

    Performed under reducing conditions.

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Product protocols

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