C5a-R/C5ar1 Antibody

SKU: orb669065

Description

Anti-C5a-R/C5ar1 Antibody. Tested in ELISA, Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Mouse.

Images & Validation

• Tested Applications: ELISA, FC, ICC, IF, WB
• Reactivity: Mouse
• Application Notes:
  Western blot, 0.25-0.5μg/ml, Mouse Immunocytochemistry/Immunofluorescence, 5μg/ml, Mouse Flow Cytometry (Fixed), 1-3μg/1x106 cells, Mouse ELISA, 0.1-0.5μg/ml, -. Add 0.2ml of distilled water will yield a concentration of 500ug/ml

Key Properties

• Antibody Type: Primary Antibody
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: Rabbit IgG
• Immunogen: E.coli-derived mouse C5a-R/C5ar1 recombinant protein (Position: H29-D327).
• Molecular Weight: 43 kDa
• Purification: Immunogen affinity purified.

Storage & Handling

• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Form/Appearance: Lyophilized
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Disclaimer: For research use only

Alternative Names

C-C chemokine receptor type 1; C-C CKR-1; CC-CKR-1; CCR-1; CCR1; Macrophage inflammatory protein 1-alpha receptor; MIP-1alpha-R; RANTES-R; CD191; Ccr1; Cmkbr1

Flow Cytometry analysis of RAW264.7 cells using anti-C5a-R/C5ar1 antibody. Overlay histogram showing RAW264.7 cells (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-C5a-R/C5ar1 Antibody (1 µg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10 µg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 µg/1x10^6) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

IF analysis of C5a-R/C5ar1 using anti-C5a-R/C5ar1 antibody. C5a-R/C5ar1 was detected in immunocytochemical section of HEPA1-6 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 µg/mL rabbit anti-C5a-R/C5ar1 Antibody overnight at 4°C. DyLight®550 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Western blot analysis of C5a-R/C5ar1 using anti-C5a-R/C5ar1 antibody. 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 50 ug of sample under reducing conditions. Lane 1: mouse spleen tissue lysates, Lane 2: mouse thymus tissue lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150 mA 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 at 0.5 µg/mL overnight at 4°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 with Tanon 5200 system. A specific band was detected for C5a-R/C5ar1 at approximately 43 KD. The expected band size for C5a-R/C5ar1 is at 43 KD.

Quick Database Links

UniProt

P30993