Anti-C5a anaphylatoxin Polyclonal Antibody 2 is a Rabbit antibody targeting C5a anaphylatoxin. Anti-C5a anaphylatoxin Polyclonal Antibody 2 can be used in IHC-P,IHC-Fr,IF,FCM.

IgG

Human

1. Blank control: HepG2.
Primary Antibody (green line): Rabbit Anti-C5a anaphylatoxin antibody (TMAB-03450)
Dilution: 2 μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-AF647
Dilution: 1 μg /test.
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 0.1% PBST for 20 min at room temperature. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
2. Paraformaldehyde-fixed, paraffin embedded (human liver); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Complement C5 beta chain) Polyclonal Antibody, Unconjugated (TMAB-03450) at 1: 200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
3. Tissue/cell: human hepatoma tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01 M, pH 6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37℃ for 20 min;
Incubation: Anti-C5a anaphylatoxin Polyclonal Antibody, Unconjugated (TMAB-03450) 1: 200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining
4. Tissue/cell: human hepatoma tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01 M, pH 6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37℃ for 20 min;
Incubation: Anti-C5a anaphylatoxin Polyclonal Antibody, Unconjugated (TMAB-03450) 1: 200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining

IHC-P: 1:100-500; IHC-Fr: 1:100-500; IF: 1:100-500; FCM: 2μg/Test

Polyclonal

KLH conjugated synthetic peptide: human C5a anaphylatoxin

Human

C5

Complement C5

Activation of C5 by a C5 convertase initiates the spontaneous assembly of the late complement components, C5-C9, into the membrane attack complex. C5b has a transient binding site for C6. The C5b-C6 complex is the foundation upon which the lytic complex is assembled.
Derived from proteolytic degradation of complement C5, C5 anaphylatoxin is a mediator of local inflammatory process. It induces the contraction of smooth muscle, increases vascular permeability and causes histamine release from mast cells and basophilic leukocytes. C5a also stimulates the locomotion of polymorphonuclear leukocytes (chemokinesis) and direct their migration toward sites of inflammation (chemotaxis).