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MMP-9 Antibody (4A3)

Catalog # NBP2-13173 | Novus Biologicals a Bio-Techne Brand

Key Product Details

Species Reactivity

Human, Mouse, Rat

Applications

ELISA, Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 kappa Clone # 4A3

Concentration

1.0 mg/ml

Product Summary for MMP-9 Antibody (4A3)

Immunogen

Synthetic peptide from the N-terminus of human MMP9 [Swiss-Prot# P14780]

Predicted Species

Canine (100%), Equine (100%), Porcine (100%), Primate (100%), Rabbit (100%). Backed by our 100% Guarantee.

Reactivity Notes

Rat reactivity reported in scientific literature (PMID:32725911). Mouse reactivity reported in scientific literature (PMID: 30772382). .

Localization

Secreted > extracellular space > extracellular matrix

Specificity

Active MMP9. It does not react with the MMP9 proenzyme or the active and proenzyme forms of MMP2.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1 kappa

Scientific Data Images for MMP-9 Antibody (4A3)

Western Blot: MMP-9 Antibody (4A3) [NBP2-13173] - Analysis of activated MMP9 expression in MMP9 proenzyme incubated with trypsin for various times.
Immunohistochemistry: MMP-9 Antibody (4A3) [NBP2-13173] - MMP-9 (green) was detected in murine brain (cortex) using MMP-9-DyLight 488 (4A3) with a concentration of 1:20 in PBS for 2 hours. MMP9-Signals are overlaped with the corresponding phase contrast image. Image from verified customer review. Image using the FITC format of this antibody.
Western Blot: MMP-9 Antibody (4A3) [NBP2-13173] - Identification of gelatinases by Western blot analysis. A pool of sera from 10 patients with inactive RR-MS was used. For 2-DZ, pooled sera were concentrated 10-fold and 40 ug of total proteins were applied (left panels). For Western blot analysis pooled sera were concentrated 40-fold and 160 ug of total proteins were subjected to 2-DE (right panels). 2-DE gels were cut and the corresponding nitrocellulose membranes were treated with anti-NGAL Ab (A'), anti-MMP-2 Ab (B') and Anti-MMP-9 (4A3) Ab, recognizing the 65 kD and the 82 kD activated forms (C'); Gel C show the region with isoelectric point between pH 4-5.5 and molecular mass between 85 and 55 kD, analysed by Western blot in C', respectively. Image collected and cropped by CiteAb from the following publication (null), licensed under a CC-BY license.

Applications for MMP-9 Antibody (4A3)

Application
Recommended Usage

ELISA

1:100 - 1:2000

Immunohistochemistry

1:100-1:200

Immunohistochemistry-Paraffin

1:100-1:200

Western Blot

1:500-1:1000
Application Notes
In Western blot a band can be seen at 82 or 63 kDa, representing the active forms of MMP9. In IHC, cytoplasmic and extracellular staining was observed in HeLa cells. Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (pH 6.0) is recommended.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 2 reviews rated 3.5 using NBP2-13173 in the following applications:

Published Applications

Read 33 publications using NBP2-13173 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

Tris-Glycine (pH 7.5), 0.15 M NaCl

Preservative

0.05% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: MMP-9

MMP9, also known as Matrix metallopeptidase 9 or gelatinase B is a member of the matrix metalloproteinase (MMP) family. The MMPs are zinc-containing enzymes involved in the degradation of the extracellular matrix in normal physiological processes including embryonic development, reproduction and tissue remodeling. Most MMP family members are secreted as inactive proproteins that become activated when cleaved by extracellular proteinases. The proenzyme MMP9 is thought to be a 92 kDa protein. Processing of the precursor MMP9 yields different active forms, two of which were identified in Duncan et al 1998 (PMID: 9851689) to be 82 kDa and 63 kDa proteins. The 82 kDa protein results from cleavage at the Arg87-Phe88 bond. And the 63 kDa protein is caused by a further cleavage to the 82 kDa protein within the C-terminus domain between Arg527-Gly528. The main 63 kDa protein produced thus has the N-terminus of activated MMP9 and has a cleaved C-terminus. Antibodies specific to the active forms of MMP9 can be useful tools for studying important pathological processes such as tumor invasion, metastasis and arthritis.

Long Name

Matrix Metalloproteinase 9

Alternate Names

CLG4B, Gelatinase B, GELB, MANDP2, MMP9

Entrez Gene IDs

4318 (Human)

Gene Symbol

MMP9

UniProt

Product Documents for MMP-9 Antibody (4A3)

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for MMP-9 Antibody (4A3)

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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Protocols

View specific protocols for MMP-9 Antibody (4A3) (NBP2-13173):

[[URL:https://www.novusbio.com/products/mmp-9-antibody-4a3_nbp2-13173]][[Capt… Antibody (4A3)]]
Immunohistochemistry-Paraffin Embedded Sections

Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes.

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in wash buffer for 5 minutes.
3. Block each section with 100-400 ul blocking solution for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4 C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul biotinylated diluted secondary antibody. Incubate 30 minutes at room temperature.
7. Remove secondary antibody solution and wash sections three times with wash buffer for 5 minutes each.
8. Add 100-400 ul Streptavidin-HRP reagent to each section and incubate for 30 minutes at room temperature.
9. Wash sections three times in wash buffer for 5 minutes each.
10. Add 100-400 ul DAB substrate to each section and monitor staining closely.
11. As soon as the sections develop, immerse slides in deionized water.
12. Counterstain sections in hematoxylin.
13. Wash sections in deionized water two times for 5 minutes each.
14. Dehydrate sections.
15. Mount coverslips.

*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.

FAQs for MMP-9 Antibody (4A3)

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  • Q: Would it be possible for you to send me your validation data on MMP9 Antibody IIA5 NBP1-40248? I am considering using it in my research project on colorectal cancer liver metastases.

    A: Our MMP9 Antibody IIA5 NBP1-40248 was validated in Western blot through external validation, and unfortunately, I do not have an images to share at the moment. NBP2-13173 is another very similar monoclonal antibody validated in human and rat for use in Western blot. We do have images from in-house testing for this product (please see the datasheet for NBP2-13173, under the images tab), so you may be more comfortable ordering this antibody for this target.

Showing  1 - 1 of 1 FAQ Showing All
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