Analysis Note

ControlPOSITIVE CONTROL: HFL-1 cells. Conditioned, serum-free medium from (TPA-treated) human fetal lung cells. Placenta.

Application

Anti-MMP-2 Proform Antibody, N-terminus, clone CA-4001 detects level of MMP-2 Proform & has been published & validated for use in WB, IH(P).

Western Blot (0.5-1.0 µg/mL for 2 hours at room temperature, non-reduced conditions. However, chicken samples react best when fully reduced. Antibody dilution buffers should be simple for best results, PBS or TBS with 0.05% tween only. Immunogen used is highly conserved between species, thus immunoabsorption to pro-MMP2 in milk may impair reactivity.

Immunohistochemistry on frozen and formalin-fixed, paraffin-embedded tissue sections: 2-10 µg/mL for 30 minutes at room temperature. {Seftor, et al 2001} and Gottsch ML et al 2002}. Antigen retrieval is optional in most cases. Antibody dilution solutions should not contain serum because of possible immunoabsorption of the antibody to proMMP2 present in the solution.

Blocking: Inhibition of MMP-2 Activity (2-4 µg/mL).



Optimal working dilutions must be determined by end user.

Immunogen

N-terminal peptide APSPIIKFPGDVAPKTDK of procollagenase IV, coupled to KLH.{Marqulies, et al 1992}

Legal Information

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

Linkage

Replaces: 04-1048

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Physical form

Purified from ascites fluid by Protein G chromatography. Liquid in 10 mM PBS, pH 7.4, with 0.2% BSA and 15 mM sodium azide

Format: Purified

Specificity

It recognizes a protein of 72 kDa which is identified as pro (latent) form (active form is 62 kDa) of matrix metalloproteinase-2 and any fragment with N-terminal (proform) domain still attached. Antibody shows no cross-reaction with pro and active forms of MMP-9. MMP-2 is synthesized as 631 amino acid proenzyme which is activated by cleavage of the first 80 amino acids. MMPs have a common mode of activation, a conserved amino acid sequence in the putative metal binding-active site region, and are inhibited by specific tissue inhibitors of metalloproteinases (TIMPs). Cellular Localization: Cytoplasmic.