PA5-13183

antibody from Invitrogen Antibodies

Targeting: MMP14 MT1-MMP

Western blot (Data presented on provider website)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Data presented on provider website)

Flow cytometry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-13183

Provider

Invitrogen Antibodies

Product name

MMP14 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

This antibody is predicted to react with bovine, mouse, porcine and rat based on sequence homology.

Reactivity

Human, Mouse, Rat

Host

Rabbit

Isotype

IgG

Vial size

400 µL

Concentration

0.38 mg/mL

Storage

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

References

Submitted references

Mesenchymal stromal cell-derived septoclasts resorb cartilage during developmental ossification and fracture healing.

Sivaraj KK, Majev PG, Jeong HW, Dharmalingam B, Zeuschner D, Schröder S, Bixel MG, Timmen M, Stange R, Adams RH
Nature communications 2022 Jan 28;13(1):571

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The Effect of Neddylation Inhibition on Inflammation-Induced MMP9 Gene Expression in Esophageal Squamous Cell Carcinoma.

Hryniewicz-Jankowska A, Wierzbicki J, Tabola R, Stach K, Sossey-Alaoui K, Augoff K
International journal of molecular sciences 2021 Feb 9;22(4)

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TNF-α promotes breast cancer cell migration and enhances the concentration of membrane-associated proteases in lipid rafts.

Wolczyk D, Zaremba-Czogalla M, Hryniewicz-Jankowska A, Tabola R, Grabowski K, Sikorski AF, Augoff K
Cellular oncology (Dordrecht) 2016 Aug;39(4):353-63

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Serous and mucinous ovarian tumors express different profiles of MMP-2, -7, -9, MT1-MMP, and TIMP-1 and -2.

Brun JL, Cortez A, Commo F, Uzan S, Rouzier R, Daraï E
International journal of oncology 2008 Dec;33(6):1239-46

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of MMP14 was performed using 70% confluent log phase MDA-MB-231 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with MMP14 Polyclonal Antibody (Product # PA5-13183) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing golgi and cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry analysis of MCF-7 cells using a MMP14 polyclonal antibody (Product # PA5-13183) (bottom), compared to a negative control cell (top) at a dilution of 1:10-50, followed by a FITC-conjugated goat anti-rabbit antibody

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 1 Effect of tumor necrosis factor-alpha (TNF-alpha) and MLN4924 on matrix metalloproteinase 9 (MMP9) expression and esophageal squamous cell carcinoma (ESCC) cell migration. ( A ) After overnight serum starvation, KYSE150 cells were pretreated or not with 1 muM MLN4924 for 30 min and then stimulated with TNF-alpha at the indicated concentration for 24 h. Activity of MMP9 and MMP2 was analyzed by gelatin zymography in the conditioned media. The protein levels of membrane type I-matrix metalloproteinase (MT1-MMP), cyclin dependent kinase inhibitor 1A (CDKN1A/p21), c-Jun, nuclear factor kappa B (NFkappaB) and SP1 were determined by Western blotting. Fold change calculated as the ratio of relative levels of proteins normalized to beta-actin (ACTB) between the cells treated with TNF-alpha in combination with MLN4924 and the cells treated with TNF-alpha alone is shown in Figure S4 . Values shown are means +- SEM: whiskers: min-max. ( B ) MMP9 messenger RNA (mRNA) expression was analyzed by reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) in both KYSE150 and KYSE70 cells treated for 24 h with TNF-alpha (30 ng/mL) and MLN4924 (1 µM) alone or in combination. Results are presented as fold change of MMP9 gene in the treated cells relative to the untreated controls normalized to the expression of the reference gene encoding glyceraldehyde 3-phosphate dehydrogenase ( GAPDH ). ( C ) Representative images from wound healing assay showing the chan