Immunofluorescence analysis of TIMP2 was done on 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with TIMP2 (F27 P3 A4) Mouse Monoclonal Antibody (MA1774) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing cytoplasmic and nuclear localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse|
|Host / Isotype||Mouse / IgG2|
|Clone||F27 P3 A4|
|Immunogen||Synthetic peptide corresponding to residues D(56) S G N D I Y G N P I(66) of mouse TIMP-2.|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||1-2 µg/ml|
|Immunofluorescence (IF)||1-2 µg/ml|
|Immunohistochemistry (IHC)||1:100 - 1:500|
|Immunohistochemistry (Paraffin) (IHC (P))||1:50-1:500|
|Western Blot (WB)||1:500|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
MA1-774 contains 100 ug of in vitro produced, protein A purified antibody (1 mg/ml) in PBS containing 1 mg/ml BSA and 0.05% sodium azide.
MA1-774 detects TIMP-2 from mouse samples.
MA1-774 has been successfully used in Western blot, IF and immunohistochemistry procedures. By Western blot, MA1-774 detects a ~18-24 kDa band representing TIMP-2 from mouse platelets. It is recommended to block with 3% BSA.
The MA1-774 immunogen is a synthetic peptide corresponding to residues D(56) S G N D I Y G N P I(66) of mouse TIMP-2. This sequence is 100% conserved in human, rat, rabbit, chicken, and bovine TIMP-2
Tissue inhibitor of metalloproteinases-1 (TIMP-1) and Tissue inhibitor of metalloproteinases-2 (TIMP-2) are inhibitory enzymes of matrix metalloproteinase family. TIMP-1 forms a complex of 1:1 stoichiometry with activated interstitial collagenase, activated stromelysin, active form of 72 kD Type IV collagenase (MMP-2) and latent and active forms 92 kD Type IV collagenase (MMP-9). TIMPs inhibit the proteolytic invasiveness of tumor cells and normal placental trophoblast cells.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.