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Anti-C5aR2 (extracellular) Antibody

Complement Component 5a Receptor 2, C5L2, C5a Anaphylatoxin Chemotactic Receptor 2, G Protein-Coupled Receptor 77, GPR77

Cat #: AAR-034
Alternative Name Complement Component 5a Receptor 2, C5L2, C5a Anaphylatoxin Chemotactic Receptor 2, G Protein-Coupled Receptor 77, GPR77
Lyophilized Powder yes
Type: Polyclonal
Host: Rabbit
Reactivity: m, r
Immunogen
  • Peptide (C)RRLLQEHYPPRLV(S)G, corresponding to amino acid residues 179 - 193 of mouse C5aR2 (Accession Q8BW93). Extracellular, 2nd loop.
Accession (Uniprot) Number Q8BW93
Gene ID 319430
Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
Homology Mouse – 14 out of 15 amino acid residues identical; Rat - 13 out of 15 amino acid residues identical. Will not recognize Human C5aR2.
Purity Affinity purified on immobilized antigen.
Form Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN3.
Isotype Rabbit IgG
Specificity Will not recognize Human C5aR2.
Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.
Reconstitution 25 µl, 50 µl or 0.2 ml double distilled water (DDW), depending on the sample size.
Antibody concentration after reconstitution 0.8 mg/ml.
Storage after reconstitution The reconstituted solution can be stored at 4°C for up to 1 week. For longer periods, small aliquots should be stored at -20°C. Avoid multiple freezing and thawing. Centrifuge all antibody preparations before use (10000 x g 5 min).
Preadsorption Control 1 µg peptide per 1 µg antibody
Standard quality control of each lot Western blot analysis.
Applications: ifc, ih, wb
May also work in: ic*, ip*
Western blot
  • Western blot analysis of rat brain lysate (lanes 1 and 3) and mouse brain lysate (lanes 2 and 4):
    Western blot analysis of rat brain lysate (lanes 1 and 3) and mouse brain lysate (lanes 2 and 4):
    1-2. Anti-C5aR2 (extracellular) Antibody (#AAR-034), (1:400).
    3-4. Anti-C5aR2 (extracellular) Antibody, preincubated with C5aR2 (extracellular) Blocking Peptide (BLP-AR034).
  • Western blot analysis of mouse J774 macrophage cell line lysate (lanes 1 and 3) and mouse M1 myeloid leukemia cell line lysate (lanes 2 and 4):
    Western blot analysis of mouse J774 macrophage cell line lysate (lanes 1 and 3) and mouse M1 myeloid leukemia cell line lysate (lanes 2 and 4):
    1-2. Anti-C5aR2 (extracellular) Antibody (#AAR-034), (1:400).
    3-4. Anti-C5aR2 (extracellular) Antibody, preincubated with C5aR2 (extracellular) Blocking Peptide (BLP-AR034).
Immunohistochemistry
  • Expression of C5aR2 in rat hippocampus.
    Expression of C5aR2 in rat hippocampus.
    Immunohistochemical staining of perfusion-fixed frozen rat brain sections with Anti-C5aR2 (extracellular) Antibody (#AAR-034), (1: 300), followed by goat anti-rabbit-AlexaFluor-488. A. Staining in the hippocampal CA1 region, showed C5aR2 immunoreactivity (green) in glial profiles (arrows). B. Pre-incubation of the antibody with C5aR2 (extracellular) Blocking Peptide (BLP-AR034), suppressed staining. Cell nuclei are stained with DAPI (blue). P = pyramidal layer, SO = stratum oriens, SR = stratum radiatum.
Indirect flow cytometry
  • Cell surface detection of C5aR2 by indirect flow cytometry in live intact mouse J774 macrophage cell line:
    Cell surface detection of C5aR2 by indirect flow cytometry in live intact mouse J774 macrophage cell line:
    ___ Cells.
    ___ Cells + goat-anti-rabbit-FITC.
    ___ Cells + Anti-C5aR2 (extracellular) Antibody (#AAR-034), (2.5μg) + goat-anti-rabbit-FITC.
References
  1. Dunkelberger,J.R and Chao Song (2009). Cell Research. 20, 34.
  2. Janeway,C.A (2001). Immunobiology, 5th edition. 10, 3642.
  3. The Complement FactsBook. The Complement FactsBook (2018). doi:10.1016/c2015-0-06595-9.
  4. Jore,M.M (2016). Nat Struct Mol Biol. 23,378.
  5. The Complement System (2014). Pathobiology of Human Disease. doi:10.1016/B978-0-12-386456-7.01802-5.
  6. Xaria X. Li (2019). J Immunol. 202, 3348.
  7. Zhang, T (2017). J Immunol. Res. 2017, 8193932.
Scientific background

The complement system is a vital component of the innate immune system that play an important role in the immune responses to infectious organisms and tissue damages 1. The activation of the complement system is carried out through a proteolytic enzymes cascade generating various peptides and fragments 2.

One of the most important component of the complement system is a protein named complement protein C5 3. Upon proteolytic cleavage of C5 two peptides are generated, C5a and C5b 4. C5a is known to be the most potent chemotactic factor in cells and responsible for the directed migration of cells involved in the immune response to infectious organisms and tissue damages 5.    

There are two different C5a receptors, C5aR1 (C5aR, CD88) and C5aR2 (C5L2, GPR77). C5aR1 is a member of the G-protein coupled receptors (GPCRs), and known to mediate the vast majority of C5a biological activity 6. In contrast C5aR2 does not exhibit any functional coupling to G proteins and it exact mechanism of action remains controversial 7

Expression of C5aR2 can be found in various tissues such as, bone marrow, spleen, lung and in immune cells such as myeloid cells and T cells 3.

Application key:

CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, IF- Immunofluorescence, IFC- Indirect flow cytometry, IHC- Immunohistochemistry, IP- Immunoprecipitation, LCI- Live cell imaging, N- Neutralization, WB- Western blot

Species reactivity key:

H- Human, M- Mouse, R- Rat
Last update: 11/04/2021

Anti-C5aR2 (extracellular) Antibody (#AAR-034) is a highly specific antibody directed against an extracellular epitope of the mouse protein. The antibody can be used in western blot, immunohistochemistry and flow cytometry applications. It has been designed to recognize C5aR2 from mouse and rat samples.

The antibody will not recognize C5aR2 from human samples.

For research purposes only, not for human use
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