Ccl3 Antibody (OASA08460)

Catalog#: OASA08460

• Size: 0.1mg

• Description of Target: RABBIT ANTI MOUSE MIP-1 ALPHA
• Gene Symbol: Ccl3
• Alias Symbols: Mip1a; Scya3; G0S19-1; AI323804; MIP1-(a); LD78alpha; MIP-1alpha; MIP1-alpha; Ccl3
• Protein Accession#: NP_035467.1
• Swissprot Id: P10855
• Host: Rabbit
• Protein Size (# AA): 92
• Specificity: MIP-1 ALPHA
• Application: ELISA, WB
• Immunogen: The immunogen for anti-Ccl3 antibody: recombinant mouse MIP-1 alpha
• Product Format: Purified IgG - lyophilised
• Isotype: Polyclonal IgG
• Applications Info: ELISA:    0.5ug/ml
  Western Blotting:    0.1ug/ml - 0.2ug/ml
  
• Additional Information: Note: For long term storage the addition of 0.09% sodium azide is recommended.
  N.B. For functional studies do not add azide.
  Preservative Stabilisers: None present
  
  Antiserum Preparation: Antisera to mouse MIP-1 alpha were raised by repeated immunisations of rabbits with highly purified antigen. Purified IgG was prepared from whole serum by affinity chromatography.
  Approx Protein Conc: IgG concentration 1.0 mg/ml
  
  Buffer Solutions: Phosphate buffered saline pH7.4
• Reconstitution and Storage: Reconstitution: Reconstitute with 0.1 ml distilled water
  Storage: Prior to reconstitution store at +4^oC. Following reconstitution store at -20^oC.
  
  This product should be stored undiluted.
  
  Storage in frost-free freezers is not recommended. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.

Product Protocol: Ccl3 antibody used to evaluate the role of macrophage inflammatory protein 1 alpha in schistosoma mansoni egg-induced granulomatous inflammation (OASA08460)

Specificity: MIP-1 ALPHA
Product Page: Ccl3 antibody (OASA08460)
Antibody Pair Available: OASA08460 Capture OASA08461 Detection
Experiment Type: MIP-1 ALPHA ELISA

Protocol:
Extracellular immunoreactive MIP1 ALPHA was quantitated using a modification of a double ligand method. Briefly, flat-bottomed 96-well microtiter plates (Immuno-Plate I 96-F; Nunc, Roskilde, Denmark) were coated with 5ul/well of rabbit anti-MIP1 ALPHA antibody (1 ng/ul in 0.6 M NaC1, 0.26 M H3BO4, and 0.08 N NaOH, pH 9.6) for 16 h at 4~ and then washed with PBS, pH 7.5, 0.05% Tween 20 (wash buffer). Microtiter plate nonspecific binding sites were blocked with 2% BSA in PBS and incubated for 90 min at 37 degree c Plates were rinsed four times with wash buffer, and diluted (1:2 and 1:10) cell-free supernatants in duplicate were added, followed by incubation for 1 h at 37 degree c Plates were washed four times, followed by the addition of 5ul/well biotinylated rabbit anti-MIP1 ALPHA antibody (3.5 ng/ul in PBS, pH 7.5, 0.05% Tween 20, and 2% FCS), and plates incubated for 30 min at 37 degree c. Plates were washed four times, streptavidin-peroxidase conjugate (Bio-Rad Laboratories, Richmond, CA) added, and the plates incubated for 30 rain at 37 degree c Plates were again washed four times and chromogen substrate (Bio-Rad Laboratories) added. The plates' reaction terminated with 50ul/well of 3 M H2SO4 solution. Plates were read at 490 nm in an ELISA reader. Standards were one-half log dilutions of recombinant MIP1 ALPHA from 1 pg/ml to 100 ng/ml. This ELISA method consistently detected MIP-la concentrations above 10 pg/ml.

Summary:
1. Treatment of mice with rabbit anti-mouse MIP-1 alpha antibodies significantly decreased 8-d primary granuloma formation (> 40%) when compared with control mice. Anti-MIP-1 alpha sera also decreased vigorous (> 20%), but not modulated granuloma formation. These findings demonstrate that MIP-1 alpha contributes to cellular recruitment during schistosome egg granuloma formation. 2. A representative photomicrograph demonstrates the abrogation due to in vivo neutralization of MIP-1 alpha in primary granuloma formation. In many lesions, nearly all of the cellular infiltration was inhibited by the anti-MIP-1 alpha treatment.

References:
1: Lukacs NW, Kunkel SL, Strieter RM, Warmington K, Chensue SW. The role of macrophage inflammatory protein 1 alpha in Schistosoma mansoni egg-induced granulomatous inflammation. J Exp Med. 1993 Jun 1;177(6):1551-9. PubMed PMID: 8496676; PubMed Central PMCID: PMC2191049.