Revision 3

#33437Store at -20C

1 Kit

(9 x 20 microliters)

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
MMP-1 (E9S9N) Rabbit mAb 54376 20 µl 55 kDa Rabbit IgG
MMP-2 (D4M2N) Rabbit mAb 40994 20 µl 64,72 kDa Rabbit IgG
MMP-9 (D6O3H) XP® Rabbit mAb 13667 20 µl 84, 92 kDa Rabbit IgG
MMP-13 (E4W3T) Rabbit mAb 69926 20 µl 60 kDa Rabbit IgG
Fibronectin/FN1 (E5H6X) Rabbit mAb 26836 20 µl 300 kDa Rabbit IgG
MMP-3 (D7F5B) Rabbit mAb 14351 20 µl 60 kDa Rabbit IgG
LOX (D8F2K) Rabbit mAb 58135 20 µl 54, 56 kDa Rabbit IgG
COL11A1 (E6O7R) Rabbit mAb 70458 20 µl 250 kDa Rabbit IgG
COL1A1 (E6A8E) Rabbit mAb 39952 20 µl 220 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The ECM Profiling Antibody Sampler Kit provides an economical means of detecting endogenous levels of the specific ECM components using the corresponding antibodies. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Background

The extracellular matrix (ECM) is a cell surface associated three-dimensional macromolecular network composed of collagens, proteoglycans/glycosaminoglycans, elastin, fibronectin, laminins, and several other glycoproteins (1). The network provides a dynamic microenvironment surrounding the cell enabling it to carry on its function. Among the ECM proteins, fibronectin functions as a mediator to bridge distinct ECM components such as collagens, growth factors, as well as cell surface integrins to regulate ECM structural change and initiate signaling pathways (2). During normal development and pathological conditions, the ECM network is highly dynamic and continuously undergoes remodeling marked by the change of the ECM structural components, such as COL1A1, COL11A1, fibronectin, and versican. The matrix-degrading enzyme MMPs such as MMP1, MMP2, and MMP9 are highly involved in this process (4). Additional players in this process are the LOX family members of lysyl oxidase. They catalyze the first step of the covalent cross-linking of ECM proteins, collagens, and elastin, which contributes to ECM stiffness and mechanical properties (5).

  1. Theocharis, A.D. et al. (2019) FEBS J 286, 2830-69.
  2. Zollinger, A.J. and Smith, M.L. (2017) Matrix Biol 60-1, 27-37.
  3. Paolillo, M. and Schinelli, S. (2019) Int J Mol Sci 20, pii: E4947. doi: 10.3390/ijms20194947.
  4. Bonnans, C. et al. (2014) Nat Rev Mol Cell Biol 15, 786-801.
  5. Amendola, P.G. et al. (2019) Cancers (Basel) 11, pii: E729. doi: 10.3390/cancers11050729.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    XP is a registered trademark of Cell Signaling Technology, Inc.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    使用に関する制限

    法的な権限を与えられたCSTの担当者が署名した書面によって別途明示的に合意された場合を除き、 CST、その関連会社または代理店が提供する製品には以下の条件が適用されます。お客様が定める条件でここに定められた条件に含まれるものを超えるもの、 または、ここに定められた条件と異なるものは、法的な権限を与えられたCSTの担当者が別途書面にて受諾した場合を除き、拒絶され、 いかなる効力も効果も有しません。

    研究専用 (For Research Use Only) またはこれに類似する表示がされた製品は、 いかなる目的についても FDA または外国もしくは国内のその他の規制機関により承認、認可または許可を受けていません。 お客様は製品を診断もしくは治療目的で使用してはならず、また、製品に表示された内容に違反する方法で使用してはなりません。 CST が販売または使用許諾する製品は、エンドユーザーであるお客様に対し、使途を研究および開発のみに限定して提供されるものです。 診断、予防もしくは治療目的で製品を使用することまたは製品を再販売 (単独であるか他の製品等の一部であるかを問いません) もしくはその他の商業的利用の目的で購入することについては、CST から別途許諾を得る必要があります。 お客様は以下の事項を遵守しなければなりません。(a) CST の製品 (単独であるか他の資材と一緒であるかを問いません) を販売、使用許諾、貸与、寄付もしくはその他の態様で第三者に譲渡したり使用させたりしてはなりません。また、商用の製品を製造するために CST の製品を使用してはなりません。(b) 複製、改変、リバースエンジニアリング、逆コンパイル、 分解または他の方法により製品の構造または技術を解明しようとしてはなりません。また、 CST の製品またはサービスと競合する製品またはサービスを開発する目的で CST の製品を使用してはなりません。(c) CST の製品の商標、商号、ロゴ、特許または著作権に関する通知または表示を除去したり改変したりしてはなりません。(d) CST の製品をCST 製品販売条件(CST’s Product Terms of Sale) および該当する書面のみに従って使用しなければなりません。(e) CST の製品に関連してお客様が使用する第三者の製品またはサービスに関する使用許諾条件、 サービス提供条件またはこれに類する合意事項を遵守しなければなりません。

    Revision 3
    #33437

    ECM Profiling Antibody Sampler Kit

    ECM Profiling Antibody Sampler Kit: Image 1 Expand Image
    Immunohistochemical analysis of paraffin-embedded SCaBER cell pellet (left, positive) or HT-29 cell pellet (right, negative) using MMP-1 (E9S9N) Rabbit mAb.
    ECM Profiling Antibody Sampler Kit: Image 2 Expand Image
    Immunohistochemical analysis of paraffin-embedded human ductal breast carcinoma (left), lung adenocarcinoma (middle) or squamous cell lung carcinoma (right) using MMP-1 (E9S9N) Rabbit mAb (top) or an MMP-1 Rabbit mAb (bottom). These two antibodies detect unique, non-overlapping epitopes on human MMP-1. The similar staining patterns obtained with both antibodies help to confirm the specificity of the staining.
    ECM Profiling Antibody Sampler Kit: Image 3 Expand Image
    Immunohistochemical analysis of paraffin-embedded human esophageal carcinoma using MMP-1 (E9S9N) Rabbit mAb.
    ECM Profiling Antibody Sampler Kit: Image 4 Expand Image
    Immunohistochemical analysis of paraffin-embedded human lung adenocarcinoma using MMP-1 (E9S9N) Rabbit mAb.
    ECM Profiling Antibody Sampler Kit: Image 5 Expand Image
    Confocal immunofluorescent analysis of SCaBER cells (left, positive) or HT-29 cells (right, negative), using MMP-1 (E9S9N) Rabbit mAb (green), DyLight 650 Phalloidin #12956 (red), and DAPI #4083 (blue).
    ECM Profiling Antibody Sampler Kit: Image 6 Expand Image
    Western blot analysis of concentrated, serum-free cultured medium from U-2 OS cells, untreated (-) or treated with TPA #4174 (200 nM, 48 hr; +), using MMP-9 (D6O3H) XP® Rabbit mAb.
    ECM Profiling Antibody Sampler Kit: Image 7 Expand Image
    Western blot analysis of extracts from various cell lines using MMP-3 (D7F5B) Rabbit mAb.
    ECM Profiling Antibody Sampler Kit: Image 8 Expand Image
    Western blot analysis of extracts from various cell lines using Fibronectin/FN1 (E5H6X) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). The absence of Fibronectin/FN1 expression in PANC-1 cell extracts is consistent with molecular and proteomic expression profiling data, confirming specificity of the antibody.
    ECM Profiling Antibody Sampler Kit: Image 9 Expand Image
    Western blot analysis of extracts from various cell lines using COL1A1 (E6A8E) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
    ECM Profiling Antibody Sampler Kit: Image 10 Expand Image
    Western blot analysis of extracts from various cell lines using MMP-2 (D4M2N) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
    ECM Profiling Antibody Sampler Kit: Image 11 Expand Image
    Western blot analysis of extracts from various cell lines using MMP-1 (E9S9N) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
    ECM Profiling Antibody Sampler Kit: Image 12 Expand Image
    Western blot analysis of extracts from various cell lines using LOX (D8F2K) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
    ECM Profiling Antibody Sampler Kit: Image 13 Expand Image
    Western blot analysis of extracts from various cell lines using MMP-13 (E4W3T) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
    ECM Profiling Antibody Sampler Kit: Image 14 Expand Image
    Western blot analysis of extracts from various cell lines using COL11A1 (E6O7R) Rabbit mAb (upper), or

    β-Actin (D6A8) Rabbit mAb #8457 (lower). As expected, A-204 cells have higher levels of COL11A1 expression as compared to the other cell lines shown.

    ECM Profiling Antibody Sampler Kit: Image 15 Expand Image
    After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
    ECM Profiling Antibody Sampler Kit: Image 16 Expand Image
    Western blot analysis of extracts and concentrated culture medium from U-2 OS cells, untreated (-) or treated with TPA #4174 (200 nM, 48 hr; +) using MMP-9 (D6O3H) XP® Rabbit mAb. MMP-9 is induced by TPA treatment as expected.
    ECM Profiling Antibody Sampler Kit: Image 17 Expand Image
    Immunoprecipitation of Fibronectin/FN1 protein from Hep G2 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Fibronectin/FN1 (E5H6X) Rabbit mAb. Western blot analysis was performed using Fibronectin/FN1 (E5H6X) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.
    ECM Profiling Antibody Sampler Kit: Image 18 Expand Image
    Immunoprecipitation of COL1A1 protein from SK-N-SH cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is COL1A1 (E6A8E) Rabbit mAb. Western blot analysis was performed using COL1A1 (E6A8E) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as secondary antibody.

    .

    ECM Profiling Antibody Sampler Kit: Image 19 Expand Image
    Immunoprecipitation of MMP-2 protein from U-118MG cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is MMP-2 (D4M2N) Rabbit mAb. Western blot analysis was performed using MMP-2 (D4M2N) Rabbit mAb as primary antibody. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as secondary antibody.
    ECM Profiling Antibody Sampler Kit: Image 20 Expand Image
    Immunoprecipitation of MMP-1 protein from SCaBER cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is MMP-1 (E9S9N) Rabbit mAb. Western blot analysis was performed using MMP-1 (E9S9N) Rabbit mAb. Anti-rabbit IgG (light chain specific), HRP-linked Antibody was used as secondary antibody.
    ECM Profiling Antibody Sampler Kit: Image 21 Expand Image
    Immunohistochemical analysis of paraffin-embedded human squamous cell lung carcinoma using MMP-1 (E9S9N) Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
    ECM Profiling Antibody Sampler Kit: Image 22 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma using MMP-9 (D6O3H) XP® Rabbit mAb.
    ECM Profiling Antibody Sampler Kit: Image 23 Expand Image
    Immunohistochemical analysis of paraffin-embedded human neuroendocrine carcinoma of the lung using Fibronectin/FN1 (E5H6X) Rabbit mAb performed on the Leica® BOND Rx.
    ECM Profiling Antibody Sampler Kit: Image 24 Expand Image
    Confocal immunofluorescent analysis of U-118 MG cells (left, positive) and HT-29 cells (right, negative) using COL1A1 (E6A8E) Rabbit mAb (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
    ECM Profiling Antibody Sampler Kit: Image 25 Expand Image
    Immunohistochemical analysis of paraffin-embedded human bladder adenocarcinoma using MMP-2 (D4M2N) Rabbit mAb.
    ECM Profiling Antibody Sampler Kit: Image 26 Expand Image
    Immunohistochemical analysis of paraffin-embedded human lung carcinoma using MMP-9 (D6O3H) XP® Rabbit mAb.
    ECM Profiling Antibody Sampler Kit: Image 27 Expand Image
    Immunohistochemical analysis of paraffin-embedded Hep G2 cell pellet (left, positive) or PANC-1 cell pellet (right, negative) using Fibronectin/FN1 (E5H6X) Rabbit mAb.
    ECM Profiling Antibody Sampler Kit: Image 28 Expand Image
    Immunohistochemical analysis of paraffin-embedded U-87 MG cell pellet (left, positive) or HCT-15 cell pellet (right, negative) using MMP-2 (D4M2N) Rabbit mAb.
    ECM Profiling Antibody Sampler Kit: Image 29 Expand Image
    Immunohistochemical analysis of paraffin-embedded U-2 OS cell pellets, untreated (left) or treated with TPA #4174 (right), using MMP-9 (D6O3H) XP® Rabbit mAb.
    ECM Profiling Antibody Sampler Kit: Image 30 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using Fibronectin/FN1 (E5H6X) Rabbit mAb.
    ECM Profiling Antibody Sampler Kit: Image 31 Expand Image
    Immunohistochemical analysis of paraffin-embedded human cervical carcinoma using MMP-2 (D4M2N) Rabbit mAb.
    ECM Profiling Antibody Sampler Kit: Image 32 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon carcinoma using MMP-9 (D6O3H) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
    ECM Profiling Antibody Sampler Kit: Image 33 Expand Image
    Immunohistochemical analysis of paraffin-embedded human ductal breast carcinoma using Fibronectin/FN1 (E5H6X) Rabbit mAb.
    ECM Profiling Antibody Sampler Kit: Image 34 Expand Image
    Immunohistochemical analysis of paraffin-embedded human urothelial carcinoma using MMP-2 (D4M2N) Rabbit mAb.
    ECM Profiling Antibody Sampler Kit: Image 35 Expand Image
    Immunohistochemical analysis of paraffin-embedded human adenoid cystic carcinoma of the trachea using Fibronectin/FN1 (E5H6X) Rabbit mAb (left) or Fibronectin Rabbit mAb (right). These two antibodies detect independent, unique epitopes on human Fibronectin/FN1 protein. The similar staining patterns obtained with both antibodies help to confirm the specificity of the staining.
    ECM Profiling Antibody Sampler Kit: Image 36 Expand Image
    Confocal immunofluorescent analysis of U-87MG cells (left, positive) and HCT-15 cells (right, negative) using MMP-2 (D4M2N) Rabbit mAb (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    ECM Profiling Antibody Sampler Kit: Image 37 Expand Image
    Flow cytometric analysis of U-2 OS cells, untreated (blue) or treated with TPA (12-O-Tetradecanoylphorbol-13-Acetate) #4174 (200 nM, 24 hr; green) using MMP-9 (D6O3H) XP® Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
    ECM Profiling Antibody Sampler Kit: Image 38 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using Fibronectin/FN1 (E5H6X) Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
    ECM Profiling Antibody Sampler Kit: Image 39 Expand Image
    Immunohistochemical analysis of paraffin-embedded human lung adenocarcinoma using Fibronectin/FN1 (E5H6X) Rabbit mAb.
    ECM Profiling Antibody Sampler Kit: Image 40 Expand Image
    Immunohistochemical analysis of paraffin-embedded human B-cell non-Hodgkin lymphoma using Fibronectin/FN1 (E5H6X) Rabbit mAb.
    ECM Profiling Antibody Sampler Kit: Image 41 Expand Image
    Confocal immunofluorescent analysis of U-118 MG cells (left, positive) and PANC-1 cells (right, negative) using Fibronectin/FN1 (E5H6X) Rabbit mAb (green). Actin filaments were labeled with DyLight 650 Phalloidin #12956 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).