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Key Product Details

Species Reactivity

Human, Mouse, Rat

Applications

ELISA, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Immunoprecipitation, Knockdown Validated, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 kappa Clone # 8B4

Format

BSA Free

Concentration

1 mg/ml

Product Summary for MMP-2 Antibody (8B4) - (Pro and Active) - BSA Free

Immunogen

Activated recombinant human MMP-2. [Uniprot: P08253]

Specificity

This is specific for pro and active MMP2.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1 kappa

Scientific Data Images for MMP-2 Antibody (8B4) - (Pro and Active) - BSA Free

Western Blot: MMP-2 Antibody (8B4) - (Pro and Active) [NB200-114] - Analysis of MMP2 expression in ProMMP2 (1), Active MMP2 (2) and human pressure ulcer biopsy (3).
Immunohistochemistry-Paraffin: MMP-2 Antibody (8B4) - (Pro and Active) [NB200-114] - Detection of MMP-2 staining in FFPE human ovary tissue. Primary antibody was used at 2.5 ug/ml with the avidin-biotin-peroxidase detection method.

Applications for MMP-2 Antibody (8B4) - (Pro and Active) - BSA Free

Application
Recommended Usage

ELISA

0.03 ug/ml

Immunocytochemistry/ Immunofluorescence

1:10-1:500

Immunohistochemistry

2 ug/ml

Immunohistochemistry-Frozen

2ug/ml

Immunohistochemistry-Paraffin

2 ug/ml

Knockdown Validated

reported in scientific literature (PMID 31439546)

Western Blot

1-2ug/ml
Application Notes
In Western blot, a band is seen at 70 and 62kDa representing the MMP2 band-pair. The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Published Applications

Read 21 publications using NB200-114 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

PBS

Format

BSA Free

Preservative

0.05% Sodium Azide

Concentration

1 mg/ml

Shipping

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

Stability & Storage

Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.

Background: MMP-2

MMP2 is a member of the Matrix Metalloproteinase (MMP) family, which are involved in extracellular matrix degradation under both normal physiological and disease processes. Specifically, MMP-2 is responsible for degrading collagens type IV, V, VII, and X and gelatin type I. Other reported functions of MMP 2 include angiogenesis, tumor invasion, tissue repair, remodeling of the vasculature, inflammation, and atherosclerotic plaque rupture. MMP-2 secretion is activated by the Ras signaling pathway. The MMP2 antibody clone MMP2/8B4 was created against activated recombinant human MMP2, and will specifcally detect both the active and inactive forms. This MMP-2 antibody has been thoroughly characterized in most common immunological applications and has been cited in a number of published journal articles.

Long Name

Matrix Metalloproteinase 2

Alternate Names

Gelatinase A, MMP2

Gene Symbol

MMP2

Product Documents for MMP-2 Antibody (8B4) - (Pro and Active) - BSA Free

Certificate of Analysis

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

Product Specific Notices for MMP-2 Antibody (8B4) - (Pro and Active) - 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.

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Protocols

View specific protocols for MMP-2 Antibody (8B4) - (Pro and Active) - BSA Free (NB200-114):

[[URL:https://www.novusbio.com/products/mmp-2-antibody-8b4_nb200-114]][[Capti… Antibody (8B4)]]
For IHC on Paraffin-Embedded Tissue Sections:
1. Deparaffinize in xylene. 3 changes at 10 min each.
2. Hydrate in 100% EtOH for 5 min (2X).
3. Quench endogenous peroxidase. 30 min in 1% H2O2 in methanol. (Make fresh each time: 6 ml 30% H2O2+ 194 ml methanol).
4. Hydrate to H2O in graded alcohols.
a. 5 min in 95%.
b. 5 min in 70%
c. 5 min in distilled H2O.
5. Perform antigen retrieval (Antigen Retrieval; Dako #S1700) with heating as described by manufacturer. Fill plastic Coplin jar with 50 ml Antigen Retrieval buffer. Add slides to jar when temperature reaches 95-99C start timing. Heat for 30 min. After heating, let slides cool in jar for 15 min.
6. Wash slides in PBS for 5 min (2x).
7. Infiltrate sections with PBS + 0.5% Triton X100 for 10 min, then rinse twice in PBS.
8. Block for 1 hr. at room temp in Blocking buffer- (PBS + 10% goat serum).
9. Aspirate blocking buffer. Apply MMP2/8B4 IgG diluted to 5 ug/ml in Blocking buffer. Incubate overnight at 4C (or >2 hr at 37C).
10. Rinse 3X in PBS for 10 min each.
11. Apply 2 degrees antibody. Dilute biotin conjugated goat anti-mouse 1:500 (Dako # E0433) in Blocking buffer.
12. Incubate for 2 hr. at room temperature.
13. Rinse 3X in PBS for 10 min each.
14. Prepare Avidin-biotin-peroxidase by combining each component at 1:50 in PBS + 0.1% Triton X100. Gently mix and react for 30 min. Prior to use, dilute the complexed mixture 1:5 with PBS + 0.1% BSA. Apply to sections and incubate for 2 hr at room temp.
15. Rinse in 3X in PBS for 15 min each.
16. Develop with DAB. Dissolved 1.5 mg diaminobenzidine-(HCl)4 in 3 ml of PBS, then add 2 ul of 30% hydrogen peroxide. Filter through a 0.2 um filter immediately before use. Develop for 15 min.
17. Rinse in water for 5 min. Counterstain for nuclei (e.g., hematoxylin) as desired.
18. Dehydrate through graded ethanol, clear in 100% ethanol and xylene (1:1 solution) and then 100% xylene. Coverslip with Permount.

FAQs for MMP-2 Antibody (8B4) - (Pro and Active) - BSA Free

Showing  1 - 2 of 2 FAQs Showing All
  • Q: I ordered the MMP-2 ms monoclonal ab 8B4, and after several tries, I cannot get it to work. I have tried in my cell lysates, concentrated conditioned media and a uterine tissue, and I am still not able to see it. Would it be possible for your company to send me a lysate that I can use as a positive control so that I can test it in something where it should be expressed? Also, are there any already known conditions for western blotting?

    A: Our lab does not have any of the positive control biopsy lysate available to send you as a control. We do offer some transfected lysates that may be of interest to you. I also have some additional information about expected samples that would express the protein. You may also find this information on UniProt useful.

  • Q: Could you guarantee NB200-113 and NB200-114 both detect human pro-form and active form MMP2 ?

    A: Yes, both of your mentioned antibodies (i.e. NB200-113 and NB200-114) detects the pro (approximately 70kD) as well as the activated forms (approximately 62kD) of MMP-2 in Western Blot. In fact, the validation image on the datasheet of #NB200-114 shows a very nice blot where these two forms are very evident and this antibody has been cited in 8 peer reviewed publications.

Showing  1 - 2 of 2 FAQs Showing All
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