PA5-16851

antibody from Invitrogen Antibodies

Targeting: MMP9 CLG4B

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Data presented on provider website)

Immunohistochemistry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-16851

Provider

Invitrogen Antibodies

Product name

Anti-MMP9 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

PA5-16851 targets MMP-9 (92kDa Collagenase IV) in IHC (P) and WB applications and shows reactivity with Guinea Pig and Human samples. The PA5-16851 immunogen is a synthetic peptide derived from near C-terminal of human MMP-9 protein.

Reactivity

Human, Guinea Pig

Host

Rabbit

Isotype

IgG

Vial size

500 µL

Concentration

Lot Dependent

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot of MMP-9 (92kDa Collagenase IV) using MMP-9 (92kDa Collagenase IV) Polyclonal Antibody (Product # PA5-16851) on HFL-1+TPA med Cells.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on conditioned media of serum starved HT-29 (Lane 1), serum starved COLO 205 (Lane 2) and serum starved KARPAS 299 (Lane 3). The blots were probed with Anti-MMP-9 Rabbit Polyclonal Antibody (Product # PA5-16851, 2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjµgate (Product # A27036, 0.4µg/mL, 1:2500 dilution). A ~ 82 kDa band corresponding to MMP-9 was observed across cell lines tested. MMP9 is secreted as a 92 kDa zymogen. Cleavage of ProMMP-9 results in the active enzyme, having a molecular weight of approximately 82 kDa. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10 % Bis-Tris gel (Product # NP0302BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane by overnight transfer method. The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Formalin-fixed, paraffin-embedded human placenta stained with MMP-9 antibody2 using peroxidase-conjugate and AEC chromogen. Note membrane staining of trophoblasts.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of MMP-9 (92kDa Collagenase IV) showing staining in the cytoplasm of paraffin-embedded human tonsil tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a MMP-9 (92kDa Collagenase IV) Rabbit Polyclonal Antibody (Product # PA5-16851) diluted in 3% BSA-PBS at a dilution of 1:100 for 1 hour at 37ºC in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of MMP-9 was performed using 70% confluent log phase MCF-7 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 MMP-9 (92 kDa Collagenase IV) Rabbit Polyclonal Antibody (Product # PA5-16851) at 2µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjµgate (Product # A27034) 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 (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.