Anti-MMP9 antibody [EPR22140-154]

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-MMP9 antibody [EPR22140-154] (AB228402)

Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen mouse lung tissue labeling MMP9 with ab228402 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1,000 dilution (green). Positive staining on sporadic cells (macrophage-like) in mouse lung tissue is observed.

The nuclear counterstain is DAPI (blue).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1,000 dilution.

Heat-mediated antigen retrieval using sodium citrate buffer (10 mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP9 antibody [EPR22140-154] (AB228402)

Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling MMP9 with ab228402 at 1/5,000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on polymorphonuclear leukocytes of mouse spleen (PMID : 28775117). Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Heat-mediated antigen retrieval using citrate buffer (pH 6.0).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP9 antibody [EPR22140-154] (AB228402)

Immunohistochemical analysis of paraffin-embedded mouse lung tissue labeling MMP9 with ab228402 at 1/5,000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on polymorphonuclear leukocytes of mouse lung (PMID : 28775117). Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Heat-mediated antigen retrieval using citrate buffer (pH 6.0).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP9 antibody [EPR22140-154] (AB228402)

Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling MMP9 with ab228402 at 1/5,000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on polymorphonuclear leukocytes of rat spleen (PMID : 28775117). Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Heat-mediated antigen retrieval using citrate buffer (pH 6.0).

• IP

Supplier Data

Immunoprecipitation - Anti-MMP9 antibody [EPR22140-154] (AB228402)

MMP9 was immunoprecipitated from 0.35 mg of mouse brain lysate with ab228402 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab228402 at 1/1,000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5,000 dilution.

Lane 1 : Mouse lung lysate 10 μg (Input).

Lane 2 : ab228402 IP in mouse lung lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab228402 in mouse lung lysate.

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : 20 seconds.

The blot was developed using a high sensitivity ECL.

All lanes:

Immunoprecipitation - Anti-MMP9 antibody [EPR22140-154] (ab228402)

Predicted band size: 78 kDa

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• WB

Supplier Data

Western blot - Anti-MMP9 antibody [EPR22140-154] (AB228402)

Exposure times : Lanes 1-2 : 3 minutes; Lane 3 : 26 seconds.

Blocking/Dilution buffer : 5% NFDM/TBST.

MMP9 is a glycoprotein (PMID : 29329315). The 92 kDa band likely represents the zymogen prior to activating cleavage of the pro-peptide (PMID : 7688350; PMID : 12901881).

All lanes:

Western blot - Anti-MMP9 antibody [EPR22140-154] (ab228402) at 1/1000 dilution

Lane 1:

Mouse lung lysate at 20 µg

Lane 2:

Rat lung lysate at 20 µg

Lane 3:

Rat spleen lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 78 kDa

Observed band size: 92 kDa

false

• WB

Lab

Western blot - Anti-MMP9 antibody [EPR22140-154] (AB228402)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. ab181602 was used as loading control for GAPDH. Although MMP9 has been studied in brain in some publications, ab228402 was unable to detect signal in normal brain tissue， this may because MMP9 expression level is low in normal brain and would be increased in abnormal conditions like injury (PMID : 31198417)

All lanes:

Western blot - Anti-MMP9 antibody [EPR22140-154] (ab228402) at 1/1000 dilution

Lane 1:

Rat lung tissue lysate at 20 µg

Lane 2:

Rat brain tissue lysate at 20 µg

Lane 3:

Rat spleen tissue lysate at 20 µg

Lane 4:

Rat kidney tissue lysate at 20 µg

Lane 5:

Rat lymph node tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 29 kDa

false

Exposure time: 3s