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MMP9 antibody - N-terminal region (ARP33090_T100)

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100 ul

Regular Price: $229.00

Special Price: $215.00

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Conjugation Options

ARP33090_T100-FITC Conjugated

ARP33090_T100-HRP Conjugated

ARP33090_T100-Biotin Conjugated

Gene Symbol:
MMP9
NCBI Gene Id:
4318
Official Gene Full Name:
Matrix metallopeptidase 9 (gelatinase B, 92kDa gelatinase, 92kDa type IV collagenase)
Protein Name:
Matrix metalloproteinase-9
Swissprot Id:
P14780
Protein Accession #:
NP_004985
Nucleotide Accession #:
NM_004994
Alias Symbols:
GELB, CLG4B, MMP-9, MANDP2
Replacement Item:
This antibody may replace item sc-10737, HPA001238
Description of Target:
Proteins of the matrix metalloproteinase (MMP) family are involved in the breakdown of extracellular matrix in normal physiological processes, such as embryonic development, reproduction, and tissue remodeling, as well as in disease processes, such as arthritis and metastasis. Most MMP's are secreted as inactive proproteins which are activated when cleaved by extracellular proteinases. The enzyme encoded by MMP9 degrades type IV and V collagens. Studies in rhesus monkeys suggest that the enzyme is involved in IL-8-induced mobilization of hematopoietic progenitor cells from bone marrow, and murine studies suggest a role in tumor-associated tissue remodeling.
Protein Size (# AA):
707
Molecular Weight:
78kDa
Host:
Rabbit
Clonality:
Polyclonal
Purification:
Protein A purified
Application:
IHC, WB
Tissue Tool:
Find tissues and cell lines supported by DNA array analysis to express MMP9.
RNA Seq:
Find tissues and cell lines supported by RNA-seq analysis to express MMP9.
Immunogen:
The immunogen is a synthetic peptide directed towards the N terminal region of human MMP9
Species Reactivity:
Cow, Dog, Guinea Pig, Horse, Human, Mouse, Rabbit, Rat, Sheep, Zebrafish
Predicted Homology Based on Immunogen Sequence:
Cow: 100%; Dog: 100%; Guinea Pig: 100%; Horse: 100%; Human: 100%; Mouse: 100%; Rabbit: 100%; Rat: 100%; Sheep: 100%; Zebrafish: 100%
Complete computational species homology data:
Anti-MMP9 (ARP33090_T100)
Peptide Sequence:
Synthetic peptide located within the following region: VAEHGDGYPFDGKDGLLAHAFPPGPGIQGDAHFDDDELWSLGKGVVVPTR
Product Format:
Liquid. Purified antibody supplied in 1x PBS buffer with 0.09% (w/v) sodium azide and 2% sucrose.
Reconstitution and Storage:
For short term use, store at 2-8C up to 1 week. For long term storage, store at -20C in small aliquots to prevent freeze-thaw cycles.
Concentration:
Batch dependent within range: 100 ul at 0.5 - 1 mg/ml
Protein Interactions:
CXCL5; PZP; MMP26; RECK; KISS1; PRSS2; EPHB2; BTC; LCN2; COL4A6; COL4A5; TIMP3; THBS2; THBS1; MMP10; MMP7; PLG; CXCL1; TFPI; CXCL8; FN1; MMP9; COL1A2; COL1A1; COL4A4; COL4A1; COL4A2; COL4A3; HAPLN1; CD44; AREG; TGFB1; CXCL6;
Blocking Peptide:
For anti-MMP9 (ARP33090_T100) antibody is Catalog # AAP33090 (Previous Catalog # AAPY00103)
Datasheets/Manuals:
Printable datasheet for anti-MMP9 (ARP33090_T100) antibody
Target Reference:
Nozell,S., et al., (2004) J. Biol. Chem. 279 (37), 38577-38589
Publications:

Gotschy, A. et al. Local arterial stiffening assessed by MRI precedes atherosclerotic plaque formation. Circ. Cardiovasc. Imaging 6, 916-23 (2013). IHC, WB, Cow, Dog, Guinea Pig, Horse, Human, Mouse, Rabbit, Rat, Sheep, Zebrafish 24100044

Li, L. et al. DLK1 promotes lung cancer cell invasion through upregulation of MMP9 expression depending on Notch signaling. PLoS One 9, e91509 (2014). IHC, WB, Cow, Dog, Guinea Pig, Horse, Human, Mouse, Rabbit, Rat, Sheep, Zebrafish 24621612

Puttabyatappa, M; Irwin, A; Martin, JD; Mesquitta, M; Veiga-Lopez, A; Padmanabhan, V; Developmental Programming: Gestational Exposure to Excess Testosterone Alters Expression of Ovarian Matrix Metalloproteases and Their Target Proteins. 1933719117697127 (2017). IHC, WB, Cow, Dog, Guinea Pig, Horse, Human, Mouse, Rabbit, Rat, Sheep, Zebrafish 28299992

Zhang, J., Jiang, W. & Zuo, Z. Pyrrolidine dithiocarbamate attenuates surgery-induced neuroinflammation and cognitive dysfunction possibly via inhibition of nuclear factor κB. Neuroscience 261, 1-10 (2014). IHC, WB, Cow, Dog, Guinea Pig, Horse, Human, Mouse, Rabbit, Rat, Sheep, Zebrafish 24365462

Deng, J., Zhang, J., Feng, C., Xiong, L. & Zuo, Z. Critical role of matrix metalloprotease-9 in chronic high fat diet-induced cerebral vascular remodelling and increase of ischaemic brain injury in mice†. Cardiovasc. Res. 103, 473-84 (2014). IHC, WB, Cow, Dog, Guinea Pig, Horse, Human, Mouse, Rabbit, Rat, Sheep, Zebrafish 24935427

Strilakou, A; Perelas, A; Lazaris, A; Papavdi, A; Karkalousos, P; Giannopoulou, I; Kriebardis, A; Panayiotides, I; Liapi, C; Immunohistochemical determination of the extracellular matrix modulation in a rat model of choline-deprived myocardium: the effects of carnitine. 30, 47-57 (2016). IHC, WB, Cow, Dog, Guinea Pig, Horse, Human, Mouse, Rabbit, Rat, Sheep, Zebrafish 26501493

Customer Reviews for MMP9 Antibody (ARP33090_T100) tested with prostate cancer in Immunohistochemistry

CAT#: ARP33090
MMP9 Antibody
MMP9 Antibody IHC
MMP9 Antibody, rat and mouse tissues

Immunohistochemistry Protocol:

Antibodies were tested in the range of 2 – 20 ug/mL on formaldehyde fixed rat and mouse tissues that were cut into 15 micron thick sections on the cryostat. Incubation with primary antibodies was done overnight at 4°C.

Tissue sections then were washed in PBS (pH7.4) 3 times 15 minutes each and then incubated for 1 hour at room temperature with fluorescent secondary antibodies (e.g. anti-rabbit Cy3, anti-rabbit Cy2) diluted according to manufacture's recommendation.

After that section were washed with PBS (pH7.4) 3 times 15 minutes each and then mounted under coverslips using MVS Pacific anti-fade mounting media (available for OEM) with or without nuclear counterstain (e.g. DAPI).

Each experiment was repeated twice.

Product Protocols: MMP-9 gene promoter and luciferase assay Protocol with MMP9 Antibody (Catalog Number: ARP33090_T100)

Product Protocols:
MMP-9 gene promoter and luciferase assay Protocol with Gene Name: MMP9 Antibody (Catalog Number: ARP33090_T100)

Catalog Number: ARP33090_T100

Citation:
1. Liau SS, Jazag A, Whang EE. HMGA1 is a determinant of cellular invasiveness and in vivo metastatic potential in pancreatic adenocarcinoma. Cancer Res. 2006 Dec 15,66(24):11613-22. PubMed PMID: 17178855.

Product Name: MMP9 antibody - N-terminal region (ARP33090_T100)

Gene Name: MMP9

Species: MiaPaCa2 and PANC1 human pancreatic ductal adenocarcinoma cells

Experiment Name: MMP-9 gene promoter and luciferase assay

Experiment Background:
1. The purpose of this study was to test the hypothesis that HMGA1 is a molecular determinant of cellular invasiveness and metastasis in pancreatic adenocarcinoma. Siong-Seng Liau, Amarsanaa Jazag, and Edward E. Whang s findings indicate that HMGA1 promotes cellular invasiveness through phosphatidylinositol-3 kinase (PI3K)/Akt-dependent modulation of matrix metalloproteinase-9 (MMP-9) activity.
2. To assess the effect of modulating HMGA1 expression on MMP-9 promoter activity, Siong-Seng Liau, Amarsanaa Jazag, and Edward E. Whang did cotransfection experiments in which MiaPaCa2 cells were transiently transfected with either shHMGA1 or pIRES-HMGA1 vector together with MMP-9 promoter reporter vector. MMP-9 promoter activities were significantly lower with HMGA1 silencing, whereas MMP-9 promoter activity levels were markedly higher with HMGA1 overexpression, compared with controls

Experimental Steps:
1. The MMP-9 promoter reporter construct was custom-synthesized by Aviva Systems Biology (San Diego, CA).
2. Tthe full-length human MMP-9 promoter region ranging from -992 to +304 bp, relative to the transcription initiation site, was amplified by PCR from human placental DNA, using proper primers designed according to the MMP-9 gene sequence. The primers were forward, 5'-GGTACCTCTTTCTGGGCTCAAGCAATC-3' and backward, 5'-CTCGAGCTAACCCTGGACACCTCTGTTC-3'.
3. The amplified promoter fragment with the addition of XhoI and KpnI restriction sites at each end was cloned into the pGL4.12 vector (Promega, Madison, WI) at the XhoI/KpnI site upstream of the firefly luciferase gene. By direct sequencing, the sequence of the cloned promoter region was confirmed.The empty pGL4.12 vector (pGL4e) was used as control.
4. Cells were plated onto a six-well cell culture plates at a concentration of 5 × 105 per well and allowed to adhere for 12 hours in serum-containing medium.
5. To assess the effects of modulating HMGA1 expression on MMP-9 promoter activity, transient transfection experiments were done on MiaPaCa2 cells. For HMGA1 suppression experiments, 5 ug shHMGA1 or control shRNA vector was cotransfected with 5 ug pGL4-MMP9 or pGL4e and 0.5 ug pRLCMV vector (Promega), which contains a cytomegalovirus (CMV) promoter upstream of a Renilla luciferase gene.
6. For HMGA1 overexpression experiments, either pIRES-HMGA1 or control vector pIRES-puro3 was used in cotransfection. After 48 hours, luciferase activity in lysates of the transfected cells was measured using the dual-luciferase reporter assay.

Product Protocols: MMP9 antibody tested with Human K562 Cells (ARP33090_T100)

Aviva Systems Biology is the original manufacturer of this MMP9 antibody (ARP33090_T100)

Click here to view the MMP9 antibody Western Blot Protocol

Product Datasheet Link: MMP9 antibody (ARP33090_T100)

WB Suggested Anti-MMP9 Antibody Titration: 1.25ug/ml
Positive Control: K562

Western Blot image:


Description of Target: Proteins of the matrix metalloproteinase (MMP) family are involved in the breakdown of extracellular matrix in normal physiological processes, such as embryonic development, reproduction, and tissue remodeling, as well as in disease processes, such as arthritis and metastasis. Most MMP's are secreted as inactive proproteins which are activated when cleaved by extracellular proteinases. The enzyme encoded by MMP9 degrades type IV and V collagens. Studies in rhesus monkeys suggest that the enzyme is involved in IL-8-induced mobilization of hematopoietic progenitor cells from bone marrow, and murine studies suggest a role in tumor-associated tissue remodeling.

Questions pertaining to this data can be directed to techsupport@avivasysbio.com

Aviva Systems Biology’s MMP9 antibody (ARP33090_T100) has been tested using other cell lysates and tissues. To obtain more data about this antibody please email us at info@avivasysbio.com.

To order by phone call us at (888) 880-0001, fax us at (858) 552-6975 or send an email to info@avivasysbio.com. Aviva manufactures this antibody so we can offer the best price. Please contact us to request pricing information.

All of Aviva’s products are guaranteed for the applications and experimental sample types mentioned in the datasheet below. Are you curious if this product will work for you? Please contact us at techsupport@avivasysbio.com

 
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Pig Intima Neoplasia in Swine AV fistula in IHC

Submitted by:

Arizona Kidney and Vascular program                                                                                                                Division of Nephrology Research Lab                                                                                                                  Tucson College of Medicine                                                                                                                                University of Arizona

1.       Target: MMP9  

2.       Species and tissue/cell type used: Pig/intima neoplasia in swine artenovenous fistula

3.       Primary antibody dilution: 1:350

4.       Secondary antibody: RTU vectastian kit PK-7200; Vector laboratories

5.       Secondary antibody dilution: Prediluted from vendor

6.       What fixation method was used? Formalin fixed paraffin embedded

7.       What antigen retrieval method was used? 0.1   Trypsin for 30 min

8.       Description of stains and counterstain?

          Visualization: ImmPACT DAB EqV SK-4103, Vector laboratories

          Counterstain : Hematoxilin QS H3404, Vector laboratories

9.       Protocol:

Procedure-Day 1:

    1.   Bake slides overnight and cool for 15 min

Deparaffinize:

    2.   Xylene  3 mins X3

Rehydrate:

    3.   100% EtOH    3 mins X3

     4.     95% EtOH       3 mins X1

     5.     70% EtOH        2 mins X1

     6.     Milli Q Water    2 mins X2

     7.     PBS                 2 mins X2

Antigen Retrieval:

    8. Trypsin retrieval - keep 30 min in 37C water bath then put in PBS

    9. Put all the slides from different containers into one container with PBS.

    10.    PBS             5 mins X2

    11.    Quench: 6 mL 30%  + 194 mL water for 15 mins (use immediately upon making solution)

    12.   PBS              2 mins X2

    13.    Mark the area of the tissue on the slide with PAP pen

Blocking (VECTASTAIN ABC Kit)

    14.    Block for 30 min with the 2.5% Horse Serum (pre-immune serum of the 2º antibody-comes with the kit) at RT in a humidity chamber

Primary antibody:

    15.     Wash the slides in PBS for 1 min and then add 1-2 drops of the primary antibody diluted in blocking buffer from Vectastain kit in ratio 1:350

Procedure- Day 2:

   1.        Rinse slides in PBS:   2 mins with the vacuum once and then transfer the slides to the slide holder and dip them in PBS solution and wash them gently by up and down for 20 times. Remove them after 2 minutes and put them back on the tray.

Secondary Antibody

   2.         Add 1-2 drops of the secondary antibody: (biotinylated, VectaStain Kit) incubate at RT for 30 mins

   3.         Rinse slides: 2 mins with the vacuum once and then transfer the slides to the slide holder and dip them in PBS solution and wash them gently by up and down for 20 times. Remove them after 2 minutes and put them back on the tray.

   4.          Add 1-2 drops of the VectaStain ABC Reagent and incubate at RT for 30 mins.

   5.          Rinse slides in PBS: Rinse slides: 2 mins with the vacuum once and then transfer the slides to the slide holder and dip them in PBS solution and wash them gently by up and down for 20 times. Remove them after 2 minutes and put them back on the tray.

Visualization (DAB Reagent bottles

  6.           Incubate with DAB working solution for exactly 1.5 minutes:

       a.      For 5 mL of DAB working solution (use immediately upon making)

                i.  2.5 mL of Reagent 1 (Chromogen)

                ii. 2.5 mL of Reagent 2 (Peroxide)

   7.          Stop reaction by dipping in a water then put in a container with water one by one

Counterstain with Hematoxylin QS

   8.           Put slides on the sink pit and apply Hematoxylin for 2 minutes then wash them quickly using water  bottle and put in water container

   9.           Rinse in running water for 2 mins

  10.          Immerse in saturated lithium carbonate for 1 min

  11.          Milli Q water 10 dips or 2 mins X1

  12.         70% EtOH     2 mins X1

  13.          95% EtOH    3 mins X1

  14.          100% EtOH  3 mins X1

  15.          Xylene          3 mins X3

  16.          Mount in Permount

  17.          Dry overnight in the fume hood

Followed by the specific secondary peroxidase-conjugated antibodies. The membrane was visualized by enhanced chemiluminescence (ECL) and developed using X-ray film.

 

 

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