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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.
metalloproteinase inhibitor 2; testicular secretory protein Li 57; TIMP2; tissue inhibitor of metalloproteinases 2
CSC-21K; D11Bwg1104e; DDC8; Timp-2; TIMP2