Name: Anti-MMP9 antibody [RM1020]
Brand: RabMAb
SKU: ab283575
Rating: 5 (1 reviews)

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Western blot - Anti-MMP9 antibody [RM1020] (ab283575)

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
Suitable for: IHC-Fr, WB, IP, Indirect ELISA, IHC-P, ICC/IF, Flow Cyt (Intra)
Knockout validated
Reacts with: Mouse, Rat, Human

Carrier Free

製品名

Anti-MMP9 antibody [RM1020]
MMP9 一次抗体製品一覧
製品の詳細

Rabbit recombinant multiclonal [RM1020] to MMP9
由来種

Rabbit
アプリケーション

適用あり: IHC-Fr, WB, IP, Indirect ELISA, IHC-P, ICC/IF, Flow Cyt (Intra)more details
種交差性

交差種: Mouse, Rat, Human
免疫原

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

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
ポジティブ･コントロール
- 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/IF: RAW 264.7 cells treated with LPS and BFA, U-2 OS cells treated with TPA . Flow Cyt (intra): U-2 OS cells treated with TPA, RAW 264.7 cells treated with LPS and BFA. IP: Mouse lung lysate
特記事項
This product is a recombinant multiclonal 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.

製品の状態

Liquid
保存方法

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
バッファー

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
精製度

Protein A purified
ポリ/モノ

Recombinant Multiclonal
クローン名

RM1020
アイソタイプ

IgG
研究分野
- Cancer
- Tumor biomarkers
- Enzymes
- MMPs
- Neuroscience
- Diseases
- Neuroscience
- Processes

Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Rabbit specific IHC polymer detection kit HRP/DAB (ab209101)
Isotype control
- Rabbit specific IHC polymer detection kit HRP/DAB (ab209101)
Related Products
- VeriBlot for IP Detection Reagent (HRP) (ab131366)

The Abpromise guarantee

Abpromise保証は、次のテスト済みアプリケーションにおけるab283575の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション	Abreviews	特記事項
IHC-Fr		1/100.
WB		1/1000. Predicted molecular weight: 78 kDa.
IP		1/30.
Indirect ELISA		Use at an assay dependent concentration.
IHC-P	(1)	1/5000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF		1/50.
Flow Cyt (Intra)		1/500.

特記事項
IHC-Fr 1/100.
WB 1/1000. Predicted molecular weight: 78 kDa.
IP 1/30.
Indirect ELISA Use at an assay dependent concentration.
IHC-P 1/5000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/50.
Flow Cyt (Intra) 1/500.

機能

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.
組織特異性

Produced by normal alveolar macrophages and granulocytes.
関連疾患

Intervertebral disc disease
Metaphyseal anadysplasia 2
配列類似性

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

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.
翻訳後修飾

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.
細胞内局在

Secreted, extracellular space, extracellular matrix.
Target information above from: UniProt accession P14780 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
参照データベース
- Entrez Gene: 4318 Human
- Entrez Gene: 17395 Mouse
- Entrez Gene: 81687 Rat
- Omim: 120361 Human
- SwissProt: P14780 Human
- SwissProt: P41245 Mouse
- SwissProt: P50282 Rat
- Unigene: 297413 Human
別名
- 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

Western blot - Anti-MMP9 antibody [RM1020] (ab283575)

All lanes : Anti-MMP9 antibody [RM1020] (ab283575) at 1/1000 dilution

Lane 1 : LoVo (Human colorectal adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : Huh7 (Human hepatocellular carcinoma epithelial cell) whole cell lysate
Lane 3 : MCF7(Human breast adenocarcinoma epithelial cell) whole cell lysate
Lane 4 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 5 : Caco-2 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate
Lane 6 : A549 (Human lung carcinoma epithelial cell) serum starved overnight whole cell lysate
Lane 7 : A549 (Human lung carcinoma epithelial cell) serum starved overnight, then treated with 100ng/ml TPA for 24 hours, then together with 1ug/ml BFA for another 3h whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

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

Exposure time: 60 seconds

The expression of MMP-9 can be stimulated by various agents, such as inflammatory cytokine, growth factor, and 12-O-tetradecanoylphorbol-13-acetate (TPA) (PMID:21047770, 28969043).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP9 antibody [RM1020] (ab283575)

Immunohistochemical analysis of paraffin-embedded human gastric carcinoma tissue labelling MMP9 with ab283575 at 1:5000 dilution (0.123 μg/ml), followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on immune cells of human gastric carcinoma is observed. The section was incubated with ab283575 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counter stained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.
Western blot - Anti-MMP9 antibody [RM1020] (ab283575)

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?

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] (ab283575)

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] (ab283575)

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] (ab283575)

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 (Intracellular) - Anti-MMP9 antibody [RM1020] (ab283575)

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] (ab283575)

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] (ab283575)

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] (ab283575)

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 (Intracellular) - Anti-MMP9 antibody [RM1020] (ab283575)

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] (ab283575)

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] (ab283575)

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] (ab283575)

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.
Indirect ELISA - Anti-MMP9 antibody [RM1020] (ab283575)

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] (ab283575)

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

SDS download

Country/Region

Language
Datasheet download

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ab283575 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab283575 は 37 報の論文で使用されています。

Liu H et al. Dual‑directional effect of vinorelbine combined with cisplatin or fluorouracil on tumor growth and metastasis in metronomic chemotherapy in breast cancer. Int J Oncol 64:N/A (2024). PubMed: 38063236
Zhang G et al. ATL1 inhibits the proliferation and invasion of trophoblast cells via inhibition of the mTOR signaling pathway. J Biochem Mol Toxicol 37:e23237 (2023). PubMed: 36193555
Zhan X et al. Elevated neutrophil extracellular traps by HBV-mediated S100A9-TLR4/RAGE-ROS cascade facilitate the growth and metastasis of hepatocellular carcinoma. Cancer Commun (Lond) 43:225-245 (2023). PubMed: 36346061
Chang J & Zhang W Remifentanil modulates the TLR4‑mediated MMP‑9/TIMP1 balance and NF‑κB/STAT3 signaling in LPS‑induced A549 cells. Exp Ther Med 25:79 (2023). PubMed: 36684659
Li X et al. RNA-seq reveals novel mechanistic targets of Livin in bladder cancer. BMC Urol 23:26 (2023). PubMed: 36855119

View all Publications for this product

レビューと Q&A

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Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Mouse Tissue sections (Spleen)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Tris-EDTA pH9

Permeabilization

Specification

Spleen

Blocking step

Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.015% · Temperature: 20°C

Fixative

Paraformaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

MISS. Amy Wyatt

Verified customer

投稿 May 03 2024

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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Key features and details

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製品の概要

製品名

製品の詳細

由来種

アプリケーション

種交差性

免疫原

ポジティブ･コントロール

特記事項

製品の特性

製品の状態

保存方法

バッファー

精製度

ポリ/モノ

クローン名

アイソタイプ

研究分野

関連製品

Compatible Secondaries

Isotype control

Related Products

アプリケーション

The Abpromise guarantee

ターゲット情報

機能

組織特異性

関連疾患

配列類似性

ドメイン

翻訳後修飾

細胞内局在

参照データベース

別名

画像

プロトコール

データシートおよび資料

SDS download

Datasheet download

参考文献 (37)

レビューと Q&A

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-MMP9 antibody [RM1020]