Anti-MMP9 antibody

• WB

Lab

Western blot - Anti-MMP9 antibody (AB38898)

This blot was produced using a 4-12% Bis-Tris gel under the MOPS buffer system. The gel was run at 200V for 60 minutes before being transferred onto a nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour before being incubated with anti-MMP9 antibody (ab38898; 2 microgram per mL) overnight at 4°C. Antibody binding was detected using infrared labelled goat anti-rabbit (green) antibody (diluted 1 : 20000) for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-MMP9 antibody (ab38898) at 2 µg/mL

Lane 1:

Western blot - Native human MMP9 protein (Proenzyme, monomer) (<a href='/en-us/products/proteins-peptides/native-human-mmp9-protein-proenzyme-monomer-ab157344'>ab157344</a>) at 0.1 µg

Lane 2:

Western blot - Recombinant Mouse MMP9 protein (<a href='/en-us/products/proteins-peptides/recombinant-mouse-mmp9-protein-ab39309'>ab39309</a>) at 0.1 µg

Predicted band size: 78 kDa

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• WB

Unknown

Western blot - Anti-MMP9 antibody (AB38898)

ab38898 detects a band at 92 Kd (pro-form) and a band at 88 Kd (active form). Mouse MMP9 is slightly larger than human MMP9, and the antibody detects a band at about 105 Kd. It is recommend to concentrate samples by Gelatin-agarose affinity chromatography prior to Western blot usage. A recommended starting concentration for Western blots is 1 : 1000 when using colorimetric substrates such as BCIP/NBT, and 1 : 5000 for chemiluminescent substrates. Higher concentration of antibody may be needed for non-human samples.

All lanes:

Western blot - Anti-MMP9 antibody (ab38898)

All lanes:

Human MMP9

Predicted band size: 78 kDa

Observed band size: 88 kDa,92 kDa

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• WB

CiteAb

Western blot - Anti-MMP9 antibody (AB38898)

Western Blotting using Anti-MMP9 antibody, ab38898. Publication image from Jiang, Q. et al., 2022, Nat Commun, 35058428. Legend direct from paper.

eWAT-secreted OPN regulates bone resorption.a Representative western blots (left) and quantification (right, n = 3 biologically independent samples) ofαv and β3 in BMSCs, BMDMs, and osteoclasts (Oc) in vitro. b Immunohistochemical staining for OPN and cathepsin K (CTSK) in serial sections of tibiae from mice fed for 12 weeks. Scale bar : 50 µm. c Representative western blots (left) and quantification (right, n = 3 biologically independent samples) of the bands of JNK and p-JNK in BMDMs, Oc, and Oc+rOPN after culture for 4 days in vitro. rOPN, 0.5 µg/ml. d Representative images of the osteo-erosion surface (top) and crystal violet staining of the unwashed Osteo Assay surface with or without rOPN treatment under osteoclastogenic induction for 12 days (bottom). Scale bar : 50 µm. e Quantification of the bone resorption area from d (n = 3 biologically independent samples). f Relative expression of Mmp9 in BMDMs with or without rOPN treatment under osteoclastogenic induction for 3 and 6 days, respectively (n = 3 biologically independent samples). g Representative western blots ofαv, β3, and MMP9 in the bone marrow of NFD- and HFD-fed mice at week 12. h Immunohistochemical staining of MMP9 from the proximal tibiae of mice after injection of saline or OPN-neutralizing antibody (Neu Ab) into bilateral eWAT for 8 weeks. Scale bar : 100 µm (the bottom row was magnified from the top row, scale bar : 50 µm). i, j Representative µCT images (i) and quantification (n = 5 biologically independent samples) (j) of proximal tibiae after mice were injected with saline or Neu Ab. Scale bar : 500 µm. k, l Representative µCT images (k) and quantification (n = 4 biologically independent samples) (l) of rBMAT of tibiae after mice were injected with saline or Neu Ab. Scale bar : 500 µm. Images are representative of 3 independent experiments. All data are presented as mean ± SD. One-way ANOVA with Tukey’s post-hoc test (a, c), two-tailed Welch’s t-test (e), by two-way ANOVA with Sidak’s post-hoc test (f), and two-way ANOVA with Tukey’s post-hoc test (j, l) were used. Source data are provided as a Source Data file. See also Supplementary Tables 7 and 8.

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• WB

CiteAb

Western blot - Anti-MMP9 antibody (AB38898)

western blotting using Anti-MMP9 antibody ab38898. Publication image and figure legend from Singla, D. K., Johnson, T. A., et al. 2019, Cells, PubMed 31600901.

Fibrosis is inhibited with ESCs or ES-Exos treatment post DIC. (A) Representative Masson's trichrome images demonstrating vascular (a-d) and interstitial fibrosis (e-h). (B) Percentage of vascular fibrosis quantified over the vessel area. (C) Quantitative analysis for intestinal fibrosis. (D) Representative blot and densitometric analysis for MMP-9. Error bars = mean ± S.E.M. * p < < 0.05 vs. control, #p < < 0.05 vs. Dox, one way ANOVA followed by a Tukey test, Western blot quantities are represented as A.U. * p < < 0.05 vs. control. #p < < 0.05 vs. Dox, $p = non significant (NS) Dox + ES-Exos vs. Dox + ESCs, scale bar = 100 µm, n = 5-6. Images taken at 20x.

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• WB

CiteAb

Western blot - Anti-MMP9 antibody (AB38898)

MMP9 western blot using Anti-MMP9 antibody ab38898. Publication image and figure legend from Peetermans, M., Vanassche, T., et al., 2014, BMC Microbiol, PubMed 25515118.

Gelatinase activation by SAK in skin tissue. A. Gelatin zymogram showing pro-MMP-2 and active MMP-2 in murine skin and subcutaneous tissue protein extract after incubation with staphylokinase (SAK) and human plasminogen (huPlg). Activation with APMA, a chemical MMP-activator, was used as a positive control. Quantitation of MMP-2 activation (active/total), data are mean ± SD from 3 independent experiments. *denotes p < < 0.05. B. Gelatin zymogram showing pro-MMP-2 and active MMP-2 in day 3 lesional skin samples. A sample from normal skin and an APMA-activated normal skin extract are included as controls. Quantitation of MMP-2 activation (active/total), data are mean ± SD from 3 independent experiments. P for trend <0.05. C. MMP-9 activation in day 3 lesional skin samples, assessed by Western blot. A sample from normal skin is included as control. Quantitation of MMP-9 activation (active/total), data are mean ± SD from 3 independent experiments. P for trend <0.05.

false

• WB

CiteAb

Western blot - Anti-MMP9 antibody (AB38898)

MMp9 western blot using Anti-MMP9 antibody ab38898.Publication image and figure legend from Cramer, E. P., Glenthøj, A., et al., 2012, PLoS One, PubMed 22737251.

Western blot of human granulocytes from peripheral blood, mouse granulocytes from the spleen, and tumor extracts.Lipocalin-2 (LCN2) is unable to form a heterodimer with MMP-9 in mouse. There is no difference in the post translational modification of LCN2 in tumors compared to myeloid cells in mouse. A, samples analyzed under non-reducing conditions. In humans the LCN2 : LCN2 and the MMP-9 : MMP-9 homodimers and the LCN2 : MMP-9 heterodimer are identified. In addition the MMP-9 monomer is seen. In contrast no heterodimer between LCN2 and MMP-9 is seen in the mouse, only the LCN2 monomer and both the monomeric and dimeric forms of MMP-9 are seen. B, samples analyzed under reducing conditions. The LCN2 : MMP-9 heterodimer as well as the LCN2 : LCN2 and MMP9 : MMP9 homodimers are no longer identifiable after breakage of disulfide bindings. Merely the LCN2 monomer and the MMP-9 monomer are identified in both humans and mouse. C, left panel, samples analyzed under non-reducing conditions. Right panel, reducing conditions. Only the LCN2 monomer and no heterodimer is identified under both conditions. D, left panel, samples analyzed under non-reducing conditions. The MMP-9 monomer is seen, and in some of the tumors even the homodimer is identified. Right panel, reducing conditions. The MMP-9 monomer is seen along with high molecular weight (HMW) bands of unknown origin. The HMW bands are seen in both PyMT, Lcn2+/+ and PyMT, Lcn2-/- mice and therefore cannot be the lipocalin-2 : MMP-9 heterodimer. All blots are shown in full-length size. Samples are loaded in the same order in C and D as in Figure 8. Abbreviations : WT and KO tumors : Tumors from PyMT, Lcn2+/+ or PyMT, Lcn2-/- mice respectively. Gr-1+ : Gr-1 positive cells purified from the spleen of two FVB/N mice.

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