RecombinantRabMAb

Recombinant Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)

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Western blot - Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)
  • Immunocytochemistry/ Immunofluorescence - Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)
  • Flow Cytometry - Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)
  • Immunocytochemistry/ Immunofluorescence - Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)
  • Immunohistochemistry (Frozen sections) - Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)
  • Flow Cytometry - Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)
  • Immunoprecipitation - Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)
  • Immunohistochemistry (Frozen sections) - Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)
  • ELISA - Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)
  • Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit recombinant multiclonal [RM1020] to MMP9 - BSA and Azide free
  • Suitable for: IHC-Fr, IHC-P, ICC, WB, IP, Competitive ELISA, Flow Cyt (Intra)
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Unconjugated

Overview

  • Product name

    Anti-MMP9 antibody [RM1020] - BSA and Azide free
    See all MMP9 primary antibodies

  • Description

    Rabbit recombinant multiclonal [RM1020] to MMP9 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-Fr, IHC-P, ICC, WB, IP, Competitive ELISA, Flow Cyt (Intra)more details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    This product was produced with the following immunogens:
    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

    Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: U-2 OS treated with TPA , RAW 264.7 treated with LPS and BFA, NR8383, NR8383 treated with LPS and BFA, and Human lung and mouse lung lysates. IHC-P: Human spleen, Human tonsil, Mouse spleen and Rat spleen tissues. IHC-Fr: Mouse lung and Rat lung tissues. ICC: RAW 264.7 cells treated with LPS and BFA, U-2 OS cells treated with TPA . Flow Cyt: U-2 OS cells treated with TPA, RAW 264.7 cells treated with LPS and BFA. IP: Mouse lung lysate

  • General notes

    ab283594 is the carrier-free version of ab283575.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab283594 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-Fr
Use at an assay dependent concentration.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Predicted molecular weight: 78 kDa.
IP
Use at an assay dependent concentration.
Competitive ELISA
Use at an assay dependent concentration.
Flow Cyt (Intra)
Use at an assay dependent concentration.
Notes
IHC-Fr
Use at an assay dependent concentration.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Predicted molecular weight: 78 kDa.
IP
Use at an assay dependent concentration.
Competitive ELISA
Use at an assay dependent concentration.
Flow Cyt (Intra)
Use at an assay dependent concentration.

Target

  • Function

    May play an essential role in local proteolysis of the extracellular matrix and in leukocyte migration. Could play a role in bone osteoclastic resorption. Cleaves KiSS1 at a Gly-
    -Leu bond. Cleaves type IV and type V collagen into large C-terminal three quarter fragments and shorter N-terminal one quarter fragments. Degrades fibronectin but not laminin or Pz-peptide.

  • Tissue specificity

    Produced by normal alveolar macrophages and granulocytes.

  • Involvement in disease

    Intervertebral disc disease
    Metaphyseal anadysplasia 2

  • Sequence similarities

    Belongs to the peptidase M10A family.
    Contains 3 fibronectin type-II domains.
    Contains 4 hemopexin repeats.

  • Domain

    The conserved cysteine present in the cysteine-switch motif binds the catalytic zinc ion, thus inhibiting the enzyme. The dissociation of the cysteine from the zinc ion upon the activation-peptide release activates the enzyme.

  • Post-translational
    modifications

    Processing of the precursor yields different active forms of 64, 67 and 82 kDa. Sequentially processing by MMP3 yields the 82 kDa matrix metalloproteinase-9.
    N- and O-glycosylated.

  • Cellular localization

    Secreted, extracellular space, extracellular matrix.
  • Information by UniProt

  • Database links

    • Alternative names

      • 82 kDa matrix metalloproteinase-9 antibody
      • 92 kDa gelatinase antibody
      • 92 kDa type IV collagenase antibody
      • CLG 4B antibody
      • CLG4B antibody
      • Collagenase Type 4 beta antibody
      • Collagenase type IV 92 KD antibody
      • EC 3.4.24.35 antibody
      • Gelatinase 92 KD antibody
      • Gelatinase B antibody
      • Gelatinase beta antibody
      • GelatinaseB antibody
      • GELB antibody
      • Macrophage gelatinase antibody
      • MANDP2 antibody
      • Matrix metallopeptidase 9 (gelatinase B, 92kDa gelatinase, 92kDa type IV collagenase) antibody
      • Matrix Metalloproteinase 9 antibody
      • MMP 9 antibody
      • MMP-9 antibody
      • MMP9 antibody
      • MMP9_HUMAN antibody
      • Type V collagenase antibody
      see all

    Images

    • Western blot - Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)
      Western blot - Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)
      All lanes : Anti-MMP9 antibody [RM1020] (ab283575) at 1/1000 dilution

      Lane 1 : U-2 OS (Human bone osteosarcoma epithelial cell) whole cell lysate
      Lane 2 : U-2 OS treated with 200nM TPA for 24h whole cell lysate
      Lane 3 : RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
      Lane 4 : RAW 264.7 treated with 100ng/ml lipopolysaccharide (LPS) for 4h, then together with 1ug/ml BFA for another 3h whole cell lysate
      Lane 5 : NR8383 (Rattus norvegicus lung macrophage (alveolar)) whole cell lysate
      Lane 6 : NR8383 treated with 100ng/ml lipopolysaccharide (LPS) for 4h, then together with 1ug/ml BFA for another 3h whole cell lysate
      Lane 7 : Human lung lysate
      Lane 8 : Mouse lung lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 78 kDa
      Observed band size: 84-92 kDa why is the actual band size different from the predicted?



      This data was developed using ab283575, the same antibody clone in a different buffer formulation. 

      Blocking and diluting buffer and concentration: 5% NFDM/TBST

      MMP9 is a glycoprotein (PMID: 29329315). The 92 kDa band likely represents the zymogen prior to activating cleavage of the pro-peptide (PMID: 7688350; PMID: 12901881).

      Exposure time: Lane 3-4: 3min ; Others: 37 seconds

       

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)

      This data was developed using ab283575, the same antibody clone in a different buffer formulation.

       

      Immunohistochemical analysis of paraffin-embedded Human spleen tissue labelling MMP9 with ab283575 at 1/5000 (0.123 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on the human spleen. The section was incubated with ab283575 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)

      This data was developed using ab283575, the same antibody clone in a different buffer formulation.

      Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labelling MMP9 with ab283575 at 1/5000 (0.123 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on the human tonsil. The section was incubated with ab283575 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

       

    • Immunocytochemistry/ Immunofluorescence - Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)
      Immunocytochemistry/ Immunofluorescence - Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)

      This data was developed using ab283575, the same antibody clone in a different buffer formulation.

      Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized U-2 OS cells labelling MMP9 with ab283575 at 1/50 (12.26 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

      Confocal image showing increased cytoplasmic staining in U-2 OS cells treated with 12-O-Tetradecanoylphorbol-13-acetate (200nM) for 24hours.

      Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

       

    • Flow Cytometry - Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)
      Flow Cytometry - Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)

      This data was developed using ab283575, the same antibody clone in a different buffer formulation.

      Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized U-2 OS (Human bone osteosarcoma epithelial cell) treated with TPA (200nM 24h) (Red) / Untreated control (Green) cells labelling MMP9 with ab283575 at 1/500 dilution (0.1ug) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat F(ab')2 Anti-Rabbit IgG(DyLight® 488, ab98507) at 1/500 dilution was used as the secondary antibody.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)

      This data was developed using ab283575, the same antibody clone in a different buffer formulation.

      Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labelling MMP9 with ab283575 at 1/5000 (0.123 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on the mouse spleen. The section was incubated with ab283575 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

       

    • Immunocytochemistry/ Immunofluorescence - Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)
      Immunocytochemistry/ Immunofluorescence - Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)

      This data was developed using ab283575, the same antibody clone in a different buffer formulation.

      Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 cells labelling MMP9 with ab283575 at 1/50 (12.26 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

      Confocal image showing increased cytoplasmic staining in RAW 264.7 cells treated with lipopolysaccharide (100 ng/ml) for 4 hours then together with Brefeldin A (1 ug/ml) for 3 hours.

      Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2ug/ml) dilution.

       

    • Immunohistochemistry (Frozen sections) - Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)
      Immunohistochemistry (Frozen sections) - Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)

      This data was developed using ab283575, the same antibody clone in a different buffer formulation.

      Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse lung (fresh) tissue labeling MMP9 with ab283575 at 1/100 (6.13 ug/ml) dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 (2 ug/ml) dilution (Green). The nuclear counterstain was DAPI (Blue).

      Positive staining on macrophages of mouse lung is observed.

      Secondary antibody control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 (2 ug/ml) dilution.

       

    • Flow Cytometry - Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)
      Flow Cytometry - Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)

      This data was developed using ab283575, the same antibody clone in a different buffer formulation.

      Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) treated with 100ng/ml LPS for 4h then together with 1ug/ml BFA for another 3h (Right) / Untreated control (Left) cells labelling MMP9 with ab283575 at 1/500 dilution (0.1ug). A Goat F(ab')2 Anti-Rabbit IgG(DyLight® 488, ab98507) at 1/500 dilution was used as the secondary antibody.

    • Immunoprecipitation - Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)
      Immunoprecipitation - Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)

      This data was developed using ab283575, the same antibody clone in a different buffer formulation.

      MMP9 was immunoprecipitated from 0.35 mg Mouse lung lysate 10 ug with ab283575 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab283575 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

      Lane 1: Mouse lung lysate 10 ug

      Lane 2: ab283575 IP in Mouse lung lysate

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab283575 in Mouse lung lysate

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 3 min

       

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)

      This data was developed using ab283575, the same antibody clone in a different buffer formulation.

      Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labelling MMP9 with ab283575 at 1/5000 (0.123 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on the rat spleen. The section was incubated with ab283575 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

       

    • Immunohistochemistry (Frozen sections) - Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)
      Immunohistochemistry (Frozen sections) - Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)

      This data was developed using ab283575, the same antibody clone in a different buffer formulation.

      Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat lung (fresh) tissue labeling MMP9 with ab283575 at 1/100 (6.13 ug/ml) dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 (2 ug/ml) dilution (Green). The nuclear counterstain was DAPI (Blue).

      Positive staining on macrophages of rat lung is observed.

      Secondary antibody control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 (2 ug/ml) dilution.

       

    • ELISA - Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)
      ELISA - Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)

      This data was developed using ab283575, the same antibody clone in a different buffer formulation.

      ELISA using ab283575 at varying antibody concentrations and antigen concentration at 1000 ng/ml. An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) (1/2500) was used as the secondary antibody.

    • Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)
      Anti-MMP9 antibody [RM1020] - BSA and Azide free (ab283594)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References (0)

    Publishing research using ab283594? Please let us know so that we can cite the reference in this datasheet.

    ab283594 has not yet been referenced specifically in any publications.

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