MMP-9 Antibody (4A3) - BSA Free

Novus Biologicals | Catalog # NBP2-13173

Novus Biologicals
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Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Mouse, Rat, Canine

Predicted:

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

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, ELISA

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, IF/IHC, Westen Blot, CyTOF-ready

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 kappa Clone # 4A3

Format

BSA Free
View all MMP-9 Primary Antibodies »

Product Specifications

Immunogen

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

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) - BSA Free

Western Blot: MMP-9 Antibody (4A3) [NBP2-13173]

Western Blot: MMP-9 Antibody (4A3) [NBP2-13173]

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]

Immunohistochemistry: MMP-9 Antibody (4A3) [NBP2-13173]

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]

Western Blot: MMP-9 Antibody (4A3) [NBP2-13173]

MMP-9-Antibody-4A3-Western-Blot-NBP2-13173-img0011.jpg
Western Blot: MMP-9 Antibody (4A3) [NBP2-13173]

Western Blot: MMP-9 Antibody (4A3) [NBP2-13173]

MMP-9-Antibody-4A3-Western-Blot-NBP2-13173-img0012.jpg
Immunohistochemistry-Paraffin: MMP-9 Antibody (4A3) [NBP2-13173]

Immunohistochemistry-Paraffin: MMP-9 Antibody (4A3) [NBP2-13173]

Immunohistochemistry-Paraffin: MMP-9 Antibody (4A3) [NBP2-13173] - Analysis of a FFPE tissue section of human esophageal cancer using MMP9 antibody (clone 4A3) at 1:200. The staining was developed with HRP-labelled secondary antibody and DAB reagent followed by hematoxylin counterstaining. This antibody generated a specific extracellular and cytoplasmic staining primarily in the cancer cells while the signal was pretty weak in the tumor stroma.
Immunohistochemistry-Paraffin: MMP-9 Antibody (4A3) [NBP2-13173]

Immunohistochemistry-Paraffin: MMP-9 Antibody (4A3) [NBP2-13173]

Immunohistochemistry-Paraffin: MMP-9 Antibody (4A3) [NBP2-13173] - Analysis of a FFPE section of human esophageal cancer using MMP9 antibody (clone 4A3) at 1:200. The staining was developed with HRP-labelled secondary antibody and DAB reagent followed by hematoxylin counterstaining. This antibody clone generated an extracellular and cytoplasmic staining mainly in the cancer cells while the signal was very weak in the stromal cells of the tumor.
MMP-9 Antibody (4A3)

Western Blot: MMP-9 Antibody (4A3) [NBP2-13173] -

Western Blot: MMP-9 Antibody (4A3) [NBP2-13173] - 2-DZ analysis & identification of MMP-9 by Western blot. Left panels (A & B): 2-D zymography (2-DZ) of serum samples from healthy control (A) & from an inactive RR-MS patient not subjected to therapy (same patient as in Fig. 2; B). For 2-DZ, aliquots of 35 μl of serum were resuspended in the rehydration solution & subjected to isoelectrofocusing (IEF; 1st dimension) on IPG Dry-Strips of 13 cm in a linear pH gradient of 4–7. After IEF, IPG strips were equilibrated & then applied for the 2nd dimension in a 8.5% (w/v) polyacrylamide gel copolymerized with 0.1% (w/v) gelatin. The isoforms & charge variants of MMP-2 & MMP-9 appear as clear spots of digestion on the dark background of the gel. Right panels (A' & B') represent Western blot analysis of the same sera shown in A & B. Aliquots of 35 μl of serum samples (instead of the usual 20 μl) were subjected to 2D electrophoresis (2-DE) by using 8.5% (w/v) polyacrylamide gels without gelatin. After transfer of the proteins, the nitrocellulose membranes were incubated with an anti MMP-9 (Ab-8) Mouse mAb (IA5) at concentration of 2.66 μg/ml. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/24616914), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
MMP-9 Antibody (4A3)

Western Blot: MMP-9 Antibody (4A3) [NBP2-13173] -

Western Blot: MMP-9 Antibody (4A3) [NBP2-13173] - 2-DZ analysis & identification of MMP-9 by Western blot. Left panels (A & B): 2-D zymography (2-DZ) of serum samples from healthy control (A) & from an inactive RR-MS patient not subjected to therapy (same patient as in Fig. 2; B). For 2-DZ, aliquots of 35 μl of serum were resuspended in the rehydration solution & subjected to isoelectrofocusing (IEF; 1st dimension) on IPG Dry-Strips of 13 cm in a linear pH gradient of 4–7. After IEF, IPG strips were equilibrated & then applied for the 2nd dimension in a 8.5% (w/v) polyacrylamide gel copolymerized with 0.1% (w/v) gelatin. The isoforms & charge variants of MMP-2 & MMP-9 appear as clear spots of digestion on the dark background of the gel. Right panels (A' & B') represent Western blot analysis of the same sera shown in A & B. Aliquots of 35 μl of serum samples (instead of the usual 20 μl) were subjected to 2D electrophoresis (2-DE) by using 8.5% (w/v) polyacrylamide gels without gelatin. After transfer of the proteins, the nitrocellulose membranes were incubated with an anti MMP-9 (Ab-8) Mouse mAb (IA5) at concentration of 2.66 μg/ml. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/24616914), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
MMP-9 Antibody (4A3) - BSA Free

Immunohistochemistry: MMP-9 Antibody (4A3) - BSA Free [NBP2-13173] -

Histological and immunohistochemistry results of periapical lesions in each group. (A) Representative histopathological images in periapical areas after root canal filling; (B) The inflammation grade evaluation of periapical lesions (n = 10); (C) Representative immunohistochemistry (IHC) staining of MMP-9 in periapical areas after root canal filling. Green boxes indicate peripheral periodontal tissue; (D) Representative immunohistochemistry (IHC) staining of TNF-alpha in periapical areas after root canal filling. Green boxes indicate peripheral periodontal tissue. ns p > 0.05, not significant, * p < 0.05, ** p < 0.01. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36361925), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for MMP-9 Antibody (4A3) - BSA Free

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.

Reviewed Applications

Read 3 reviews rated 3.7 using NBP2-13173 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% 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

P14780

Additional MMP-9 Products

Product Documents for MMP-9 Antibody (4A3) - BSA Free

Certificate of Analysis

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

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

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.

Citations for MMP-9 Antibody (4A3) - BSA Free

Customer Reviews for MMP-9 Antibody (4A3) - BSA Free (3)

3.7 out of 5
3 Customer Ratings
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Customer Images

Showing  1 - 3 of 3 reviews Showing All
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  • MMP-9 Antibody (4A3)
    Name: Shinford Sun
    Application: Western Blot
    Sample Tested: Human lung tumor
    Species: Human
    Verified Customer | Posted 05/24/2024
    We have ordered a variety of antibodies from the MMP family. Not all of them can be clearly detected because MMPs are secreted proteins. The antibody for MMP9 works reliably, and we recommend using it at a concentration of 1:1000.
    MMP-9 Antibody (4A3) - BSA Free NBP2-13173
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: rat smooth muscle
    Species: Rat
    Verified Customer | Posted 07/20/2016
    intestinal smooth muscle cells
    MMP-9 Antibody (4A3) - BSA Free NBP2-13173
  • Name: Kristina Knubel
    Application: Western Blot
    Sample Tested: U251 glioma cell line; whole cell lysate
    Species: Human
    Verified Customer | Posted 02/25/2014
    MMP9 in response to foretinib treatment in U251 glioma cells
    MMP-9 Antibody (4A3) - BSA Free NBP2-13173

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Protocols

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

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 (keep slides in the sodium citrate buffer at all times).

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in PBS for 5 minutes.
3. Block each section with 100-400 ul blocking solution (1% BSA in PBS) 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 HRP polymer conjugated secondary antibody. Incubate 30 minutes at room temperature.
7. Wash sections three times in wash buffer for 5 minutes each.
8. Add 100-400 ul DAB substrate to each section and monitor staining closely.
9. As soon as the sections develop, immerse slides in deionized water.
10. Counterstain sections in hematoxylin.
11. Wash sections in deionized water two times for 5 minutes each.
12. Dehydrate sections.
13. Mount coverslips.

Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 10-25 ug of total protein per lane.
2. Transfer proteins to PVDF membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain the membrane with Ponceau S (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot TBS -0.05% Tween 20 (TBST).
5. Block the membrane in 5% Non-fat milk in TBST (blocking buffer) for at least 1 hour.
6. Wash the membrane in TBST three times for 10 minutes each.
7. Dilute primary antibody in blocking buffer and incubate overnight at 4C with gentle rocking.
8. Wash the membrane in TBST three times for 10 minutes each.
9. Incubate the membrane in diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturer's instructions) for 1 hour at room temperature.
10. Wash the blot in TBST three times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturer's instructions.

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for MMP-9 Antibody (4A3) - BSA Free

Showing  1 - 2 of 2 FAQs Showing All

  • Q:  I’m looking for a pair of antibodies to MMP-9 that can be used in a sandwich assay. Do you carry any? 

    A:

    We have 10 primary antibodies for MMP-9 that have been tested in ELISA, seen here.
    It looks like 2 have been tested for capture (please note the tested species for each of these), seen here.
    6 have been tested for detection, seen here.

  • 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.

  • Q:  I’m looking for a pair of antibodies to MMP-9 that can be used in a sandwich assay. Do you carry any? 

    A:

    We have 10 primary antibodies for MMP-9 that have been tested in ELISA, seen here.
    It looks like 2 have been tested for capture (please note the tested species for each of these), seen here.
    6 have been tested for detection, seen here.

  • 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.

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