Antibody Type: Rabbit Polyclonal

Application: WB,IHC-P,IHC-Fr,ICC/IF,IF,FCM

Reactivity: Human,Mouse,Rat,Rabbit (predicted:Pig,Cow,Horse,Sheep)

IgG

Human,Mouse,Rat,Rabbit (predicted:Pig,Cow,Horse,Sheep)

1. Tissue/cell: rat brain tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37°C for 20 min;
Incubation: Anti-MMP-2 Polyclonal Antibody, Unconjugated (TMAB-01154) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
2. Tissue/cell: rat carotid artery; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37°C for 20 min;
Incubation: Anti-MMP-2 Polyclonal Antibody, Unconjugated (TMAB-01154) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
3. Paraformaldehyde-fixed, paraffin embedded (Rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (MMP2) Polyclonal Antibody, Unconjugated (TMAB-01154) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
4. Blank control (blue line): Hela (blue).
Primary Antibody (green line): Rabbit Anti-MMP2 antibody (TMAB-01154)
Dilution: 1 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC
Dilution: 1 μg/test.
Protocol
The cells were fixed with 80% methanol (5 min at-20°C) and then permeabilized with 0.1% PBS-Tween for 20 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1 X PBS/2% BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature.
5. Paraformaldehyde-fixed, paraffin embedded (Mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (MMP2) Polyclonal Antibody, Unconjugated (TMAB-01154) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
6. Sample:
SY5Y (Human) Cell Lysate at 30 μg
U2os (Human) Cell Lysate at 30 μg
U251 (Human) Cell Lysate at 30 μg
Siha (Human) Cell Lysate at 30 μg
Primary: Anti-MMP-2 (TMAB-01154) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 62 kDa
Observed band size: 60 kDa
7. Sample:
A431 (Human) Cell Lysate at 30 μg
Mcf-7 (Human) Cell Lysate at 30 μg
Primary: Anti-MMP-2 (TMAB-01154) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 62 kDa
Observed band size: 62 kDa
8. Tissue/cell: U-87MG cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (MMP2) polyclonal Antibody, Unconjugated (TMAB-01154) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nucleus.
9. Paraformaldehyde-fixed, paraffin embedded Human Glioma; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with MMP2 Polyclonal Antibody, Unconjugated (TMAB-01154) at 1:400 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
10. Paraformaldehyde-fixed, paraffin embedded Mouse Placenta; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with MMP2 Polyclonal Antibody, Unconjugated (TMAB-01154) at 1:400 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
11. Blank control (blue line): Hela (fixed with 80% methanol (5 min at-20°C) and then permeabilized with 0.1% PBS-Tween for 20 min at room temperature).
Primary Antibody (green line): Rabbit Anti-MMP2 antibody (TMAB-01154), Dilution: 1 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC, Dilution: 1 μg/test.

WB,IHC-P,IHC-Fr,ICC/IF,IF,FCM

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; ICC/IF: 1:100; IF: 1:100-500; FCM: 1μg/Test

Polyclonal

KLH conjugated synthetic peptide: human MMP2

Human

4313

Response to hypoxia,MMPs,MMPs,MMP,MMPs,Hypoxia,Cancer