Western Blot (WB)
(Figure 1. Western blot analysis of C5a-R/C5ar1 using anti-C5a-R/C5ar1 antibody (MBS1753473).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: mouse spleen tissue lysatesLane 2: mouse thymus tissue lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with rabbit anti-C5a-R/C5ar1 antigen affinity purified polyclonal antibody (Catalog # MBS1753473) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for C5a-R/C5ar1 at approximately 43KD. The expected band size for C5a-R/C5ar1 is at 43KD. )
Rabbit anti-Mouse C5a-R/C5ar1 Polyclonal Antibody | anti-C5ar1 antibody
Anti-C5a-R/C5ar1 Antibody
ICC/IF: 5ug/ml|Mouse|
FC/FACS/FCM: 1-3ug/1x106 cells|Mouse|
Direct ELISA: 0.1-0.5ug/ml|Mouse|
Western Blot (WB)
(Figure 1. Western blot analysis of C5a-R/C5ar1 using anti-C5a-R/C5ar1 antibody (MBS1753473).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: mouse spleen tissue lysatesLane 2: mouse thymus tissue lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with rabbit anti-C5a-R/C5ar1 antigen affinity purified polyclonal antibody (Catalog # MBS1753473) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for C5a-R/C5ar1 at approximately 43KD. The expected band size for C5a-R/C5ar1 is at 43KD. )
Western Blot (WB)
(Figure 1. Western blot analysis of C5a-R/C5ar1 using anti-C5a-R/C5ar1 antibody (MBS1753473).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: mouse spleen tissue lysatesLane 2: mouse thymus tissue lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with rabbit anti-C5a-R/C5ar1 antigen affinity purified polyclonal antibody (Catalog # MBS1753473) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for C5a-R/C5ar1 at approximately 43KD. The expected band size for C5a-R/C5ar1 is at 43KD. )
Immunofluorescence (IF)
(Figure 2. IF analysis of C5a-R/C5ar1 using anti-C5a-R/C5ar1 antibody (MBS1753473).C5a-R/C5ar1 was detected in immunocytochemical section of HEPA1-6 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (MBS176582) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti-C5a-R/C5ar1 Antibody (MBS1753473) overnight at 4 degree C. DyLight®550 Conjugated Goat Anti-Rabbit IgG (BA1135) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used. )
Immunofluorescence (IF)
(Figure 2. IF analysis of C5a-R/C5ar1 using anti-C5a-R/C5ar1 antibody (MBS1753473).C5a-R/C5ar1 was detected in immunocytochemical section of HEPA1-6 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (MBS176582) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti-C5a-R/C5ar1 Antibody (MBS1753473) overnight at 4 degree C. DyLight®550 Conjugated Goat Anti-Rabbit IgG (BA1135) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used. )
Flow Cytometry (FC/FACS)
(Figure 3. Flow Cytometry analysis of RAW264. 7 cells using anti-C5a-R/C5ar1 antibody (MBS1753473).Overlay histogram showing RAW264. 7 cells stained with MBS1753473 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-C5a-R/C5ar1 Antibody (MBS1753473,1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
Flow Cytometry (FC/FACS)
(Figure 3. Flow Cytometry analysis of RAW264. 7 cells using anti-C5a-R/C5ar1 antibody (MBS1753473).Overlay histogram showing RAW264. 7 cells stained with MBS1753473 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-C5a-R/C5ar1 Antibody (MBS1753473,1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
2. Atkinson, J. P. Complement system on the attack in autoimmunity. (Commentary) J. Clin. Invest. 112: 1639-1641, 2003.
3. Bao, L., Gerard, N. P., Eddy, R. L., Jr., Shows, T. B., Gerard, C. Mapping of genes for the human C5a receptor (C5AR), human FMLP receptor (FPR), and two FMLP receptor homologue orphan receptors (FPRH1, FPRH2) to chromosome 19. Genomics 13: 437-440, 1992.
NCBI and Uniprot Product Information
Uniprot Description
C5aR: a family 1 G-protein coupled receptor that stimulates the GTPase activity of Gi2. Receptor for the chemotactic and inflammatory complement factor C5a. Stimulates chemotaxis, granule enzyme release and superoxide anion production. Phosphorylated in response to C5a or after stimulation of cells with phorbol esters. Expressed on monocytes, granulocytes, dendritic cells, astrocytes and microglia.
Protein type: Membrane protein, multi-pass; Membrane protein, integral; Receptor, GPCR; GPCR, family 1
Cellular Component: cell surface; membrane; basolateral plasma membrane; apical part of cell; plasma membrane; integral to membrane; cytoplasmic vesicle; cytosol
Molecular Function: G-protein coupled receptor activity; complement component C5a receptor activity; complement component C5a binding; signal transducer activity; anaphylatoxin receptor activity; C5a anaphylatoxin receptor activity
Biological Process: neutrophil chemotaxis; apoptosis; complement receptor mediated signaling pathway; signal transduction; chemotaxis; cell proliferation in hindbrain; mRNA transcription from RNA polymerase II promoter; G-protein coupled receptor protein signaling pathway; defense response to Gram-positive bacterium; positive regulation of angiogenesis; elevation of cytosolic calcium ion concentration; response to peptidoglycan; negative regulation of neuron apoptosis; positive regulation of epithelial cell proliferation
Research Articles on C5ar1
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Product Notes
The C5ar1 c5ar1 (Catalog #AAA1753473) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-C5a-R/C5ar1 Antibody reacts with Mouse and may cross-react with other species as described in the data sheet. AAA Biotech's C5a-R/C5ar1 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunocytochemistry (ICC), Immunofluorescence (IF), Flow Cytometry (FC/FACS/FCM), Direct ELISA (EIA). WB: 0.25-0.5ug/ml|Mouse| ICC/IF: 5ug/ml|Mouse| FC/FACS/FCM: 1-3ug/1x106 cells|Mouse| Direct ELISA: 0.1-0.5ug/ml|Mouse|. Researchers should empirically determine the suitability of the C5ar1 c5ar1 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "C5a-R/C5ar1, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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