Complement C5 Antibody (PSH04-01)

Western Blot: Complement C5 Antibody (PSH04-01) [NBP3-32209] - Western blot analysis of Complement C5 on different lysates with Rabbit anti-Complement C5 antibody (NBP3-32209) at 1/2,000 dilution. Lane 1: HepG2 cell lysate (20 ug/Lane) Lane 2: Human liver tissue lysate (40 ug/Lane) Predicted band size: 188 kDa Observed band size: 188/72 kDa Exposure time: 43 seconds; ECL 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (NBP3-32209) at 1/2,000 dilution was used in 5% NFDM/TBST at 4C overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.

Immunohistochemistry: Complement C5 Antibody (PSH04-01) [NBP3-32209] - Immunohistochemical analysis of paraffin-embedded human breast tissue with Rabbit anti-Complement C5 antibody (NBP3-32209) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (NBP3-32209) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunocytochemistry/ Immunofluorescence: Complement C5 Antibody (PSH04-01) [NBP3-32209] - Immunocytochemistry analysis of HepG2 cells labeling Complement C5 with Rabbit anti-Complement C5 antibody (NBP3-32209) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Complement C5 antibody (NBP3-32209) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.

Flow Cytometry: Complement C5 Antibody (PSH04-01) [NBP3-32209] - Flow cytometric analysis of HepG2 cells labeling Complement C5. Cells were fixed and permeabilized. Then stained with the primary antibody (NBP3-32209, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

• Reactivity: Hu
• Applications: WB, Flow, ICC/IF, IHC
• Clone: PSH04-01
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1 mg/ml

Complement C5 Antibody (PSH04-01) Summary

• Additional Information: Recombinant Monoclonal Antibody
• Immunogen: Recombinant protein within human Complement C5 aa 1-900 / 1,676. (Uniprot: P01031)
• Localization: Secreted.
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Gene: C5
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • Flow Cytometry 1:1000
  • Immunocytochemistry/ Immunofluorescence 1:100
  • Immunohistochemistry
  • Immunohistochemistry-Paraffin 1:200
  • Western Blot 1:2000
• Theoretical MW: 188 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS (pH7.4), 0.1% BSA and 40% Glycerol
• Preservative: 0.05% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Protein A purified

Alternate Names for Complement C5 Antibody (PSH04-01)

• anaphylatoxin C5a analog
• C3 and PZP-like alpha-2-macroglobulin domain-containing protein 4
• C5
• complement C5
• complement component 5
• CPAMD4MGC142298
• FLJ17816
• FLJ17822

Background

C5 is synthesised in the liver as a single polypeptide chain. Before secretion the molecule is glycosylated and secreted into plasma as a 190kDa glycoprotein consisting of a disulphide linked alpha-chain (115kDa) and beta-chain (75kDa). C5 occurs in plasma at about 75mg/L. It is activated via either the classical or the alternative complement pathway, by protoelytic cleavage of C5a (11.5kDa) from the N-terminal of the alpha-chain. C5a is a potent anaphylotoxin and stimulates directed migration of neutrophils, eosinophils, basophils and monocytes. The other fragment, C5b,initiates the assembly of the membrane attack complex that mediates the cytolysis of viral and bacterial pathogens.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Gene Symbol: C5