Complement C3 Antibody (11H9)

Flow (Intracellular): Complement C3 Antibody (11H9) [NB200-540] - An intracellular stain was performed on RAW 246.7 cells with Complement C3 (11H9-3-2) antibody NB200-540 (blue) and a matched isotype control NBP2-31382 (orange). Cells were either treated with 3uM Monensin for 3 hours to block the secretion of Complement C3 (B) or grown in normal media (A). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 2 ug/mL for 30 minutes at room temperature, followed by mouse F(ab)2 IgG (H+L) PE-conjugated secondary antibody (F0102B, R&D Systems).

Immunocytochemistry/Immunofluorescence: Complement C3 Antibody (11H9) [NB200-540] - C3 protein fragments deposited on kidney cells of MPL-lpr mouse. Staining with antibody 11H9. Glomerular staining pattern.

Immunohistochemistry-Paraffin: Complement C3 Antibody (11H9) [NB200-540] - IHC-P detection of Complement C3 protein in a formalin fixed paraffin embedded tissue section of mouse liver using 1 : 100 dilution of Complement C3 antibody (clone 11H9) NB200-540. Weak but distinct membrane-cytoplasmic immunopositivity was observed in hepatocytes and few cells developed punctate membrane staining.

Immunohistochemistry-Paraffin: Complement C3 Antibody (11H9) [NB200-540] - IHC-P detection of Complement C3 protein in a formalin fixed paraffin embedded tissue section of mouse lymph node using 1 : 100 dilution of Complement C3 antibody (clone 11H9) NB200-540. This representative photomicrograph shows a membrane-cytoplasmic immunopositivity in non-germinal center cells, and few cells developed an intense staining for this target protein.

• Reactivity: Mu
• Applications: WB, Flow, Flow-IC, IA, ICC/IF, IHC, IHC-Fr, IHC-P, IP, CyTOF-ready
• Clone: 11H9
• Clonality: Monoclonal
• Host: Rat
• Conjugate: Unconjugated
• Concentration: 1.0 mg/ml

Complement C3 Antibody (11H9) Summary

• Immunogen: C57BL/6 thymocytes saturated with rat anti-Thy-1 monoclonal antibody of IgG2b subclass (RmT1)
• Localization: Secreted
• Specificity: Mouse Complement C3 and its activation products, C3b, iC3b, C3d and C3dg
• Isotype: IgG2a
• Clonality: Monoclonal
• Host: Rat
• Gene: C3
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • Western Blot 1:100-1:2000
  • Flow Cytometry 1-2 ug/mL
  • Immunoassay 0.5 ug/well in PBS
  • Immunocytochemistry/Immunofluorescence 1:10-1:500
  • Immunohistochemistry 1:10-1:500
  • Immunohistochemistry-Frozen 1:10-1:500
  • Immunohistochemistry-Paraffin 1:10-1:500
  • Immunoprecipitation 1:10-1:500
  • CyTOF-ready
  • Flow (Intracellular)
• Application Notes: For IHC-F: Tissue sections were fixed in acetone. Antigen retrieval was performed using 50% formic acid for 5 min or microwaved (700 W) in antigen unmasking solution. For IHC-P: free-floating brain tissue pretreated with 50% formic acid for 5 min. or microwaved (700W) in antigen unmasking solution for 2-5min. Blocking endogenous persoxidase by treatment with 3% H2O2 in pBS 20 min at 25 degrees C. Blocking 2hrs, 2% BSA in PBS. For IF: Fixation in 4% paraformaldehyde in PBS pH 7.4. Vibratome sections of 4 um. Pretreated with 3% hydrogen peroxide for 20 min to quench endogenous peroxidases. Microwaved in antigen unmasking solution for 2-5 minutes as antigen retrieval.

Reactivity Notes

Mouse. Not yet tested in other species.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.05% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Protein G purified

Alternate Names for Complement C3 Antibody (11H9)

• AHUS5
• ARMD9
• ASP
• C3 and PZP-like alpha-2-macroglobulin domain-containing protein 1
• complement C3
• complement component 3
• complement component C3
• CPAMD1acylation-stimulating protein cleavage product

Background

Complement component 3, or C3, is one of nine major complement proteins in the complement system. This system is vital to the immune system because of its role in killing unwanted bacteria and its part in the initiation of inflammation. The complement system is made up of a collection of proteins found in the bloodstream. C3 happens to be the most important protein as well as the most plentiful among the other proteins in the complement system. Levels of C3 in the blood are often tested in patients with autoimmune disorders to monitor the effects of their treatment. A C3 deficiency can cause increased risk for bacterial infection.

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.

Publications for Complement C3 Antibody (NB200-540)(1)

Publication using NB200-540

• Maehara N, Arai S, Mori M et al. Circulating AIM Prevents Hepatocellular Carcinoma through Complement Activation. Cell Rep. 2014 Oct 09 [PMID: 25284781] (IHC, Mouse)

Review for Complement C3 Antibody (NB200-540) (1)

Average Rating: 4

(Based on 1 review)

We have 1 review tested in 1 species: Mouse.

reviewed by: Anonymous ICC Mouse 04/01/2017

Summary

• Application: Immunocytochemistry
• Sample Tested: brain and spinal cord
• Species: Mouse

FAQs for Complement C3 Antibody (NB200-540). (Showing 1 - 1 of 1 FAQ).

1. I am trying to establish a method to measure mice C3 levels by nephelometry. I would be most grateful if you could provide me with some help, regarding the choice of the Ab. I totally understand that since such a method has never been tried, I do not expect any guaranties. 
   • We have never performed nephelometry in our lab, and do not have a protocol or advice to provide about this application. However, it seems to me that you should choose an antibody that is capable of recognizing its target in its folded conformation. Therefore, I would suggest trying an antibody that has been validated for ICC or IHC.

Bioinformatics

• Gene Symbol: C3
• Entrez:
  • 12266(Mouse)
• Uniprot:
  • P01027()