MA5-13688

antibody from Invitrogen Antibodies

Targeting: TIMP1 CLGI, EPO, TIMP

Provider product page for MA5-13688

Western blot

Immunocytochemistry

Immunohistochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [45]
Comments [0]
Validations
Western blot [2]
Immunocytochemistry [2]
Immunohistochemistry [2]
Other assay [7]

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Product number: MA5-13688 - Provider product page
Provider: Invitrogen Antibodies
Product name: TIMP1 Monoclonal Antibody (102D1)
Antibody type: Monoclonal
Antigen: Recombinant full-length protein
Description: MA5-13688 targets TIMP-1 in IHC (P) and WB applications and shows reactivity with Human samples.
Antibody clone number: 102D1
Concentration: 0.2 mg/mL

TIMP1 protein structure - MA5-13688 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on 30 µg lysate of Mouse Liver (Lane 1), membrane enriched extract MDA-MB-231 (Lane 2) and conditioned media of MDA-MB -231 (Lane 3). The blots were probed with Anti-TIMP-1 Mouse Monoclonal Antibody (Product # MA5-13688, 1-2 µg/mL) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Secondary Antibody, HRP conjugate (Product # 62-6520, 1:4000 dilution). A ~ 23 kDa band corresponding to TIMP-1 was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10 % Bis-Tris gel (Product # NP0302BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-TIMP1 Monoclonal Antibody (102D1) (Product # MA5-13688) and a 28kDa band corresponding to TIMP1 was observed in MDA-MB-231 but was absent in Daudi which is reported to be negative. An additional band at ~40kDa was observed in Daudi. Whole cell extracts (30 µg lysate) of MDA-MB-231 (Lane 1), MDA-MB-231 treated with Protein Transport Inhibitor (PTI; 1x for 4 hours) (Lane 2), Daudi (Lane 3) and Daudi treated with Protein Transport Inhibitor (1x for 4 hours) (Lane 4) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:100 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of TIMP1 was performed using 70% confluent log phase MDA-MB-231 cells treated with Protein Transport Inhibitor (PTI; 1x for 4 hours). The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with TIMP1 Monoclonal Antibody (Product # MA5-13688) at 3µg/mL dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32766) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing increased TIMP1 expression and localization to golgi and cytoplasm upon treatment with PTI. Panel e shows untreated cells with lower expression of TIMP1. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of TIMP1 was performed using 70% confluent log phase MDA-MB-231 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with TIMP-1 (102D1) Mouse Monoclonal Antibody (Product # MA5-13688) at 2 µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 1 Left: examples of TMAs with immunostaining for each protein. 200 x Right: ( A ) immunohistochemical staining of MMP2 in epithelial cells, ( B ) TIMP3 in epithelial cells and fibroblastic cells, ( C ) TIMP3 in inflammatory mononuclear cells, and ( D ) TIMP2 in epithelial cells, fibroblast and inflammatory mononuclear cells. 400 x .

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 1 Left: examples of tissue with immunostaining for the more significant proteins in patients with tumours belong to poor prognostic group (cluster 2 group: A , B and C ) and better prognostic group (cluster 1 group: D , E and F ; magnification x 400). ( A ) Immunohistochemical staining of matrix metalloproteinase (MMP)-11 in inflammatory mononuclear cells, tumour cells and fibroblastic cells. ( B ) Immunohistochemical staining of MMP-9 in inflammatory mononuclear cells, tumour cells and fibroblastic cell negatives. ( C ) Immunohistochemical staining of tissular inhibitors of metalloproteinase (TIMP)-1 in inflammatory mononuclear cells and in fibroblastic cells, tumour cell negatives. ( D ) No immunohistochemical staining of MMP-9 in inflammatory mononuclear cells, tumour cells and fibroblastic cells. ( E ) Immunohistochemical staining of MMP-11 in tumour cells, inflammatory mononuclear cells and fibroblastic cell negatives. ( F ) Immunohistochemical staining of TIMP-1 in fibroblastic cells, inflammatory mononuclear cells and tumour cell negatives. In all cases, inflammatory mononuclear cells are indicated with small arrows, and fibroblastic cells with large arrows.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 3 Molecular markers of kidney injury. a Immunohistochemical staining showing expression of injury markers KIM-1 and TIMP-1. Scale bar represents 100 mum. b Western blot for KIM-1, NGAL, TIMP-1, and beta-actin (left), and their respective quantification (right). c Quantitative real-time PCR for Kim1 , Ngal , and Timp1 in the kidney tissue. d ELISA for KIM-1, NGAL, and TIMP-1 measured in the urine. Each group has n = 5 mice. Significant difference a p < 0.05: relative to untreated control; b p < 0.05: relative to AKI; c p < 0.05: relative to AKI-EV; d p < 0.05: relative to AKI-EV-pFUS

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 7 Expression Changes in MMP-2, MMP-9, TIMP1, DDR2 and (p)SMAD2/3/total SMAD2/3 in B6 and EphA2-R-M Mouse Hearts. MMP-9, MMP-2 and TIMP-1 expression in ephrinA1-Fc-treated +/-TGF-beta primary cardiac fibroblasts are modulated in a dose-dependent manner ( n = 3/group; significance: * vs. control, + vs. 0.1 uM EA1, ++ vs. TGF-beta). Western blots of DDR2 and (p)SMAD2/3/totalSMAD2/3 show dose-dependent changes in response to ephrinA1-Fc-treatment +/-TGF-beta (NS).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 2 MK2 deficiency halts the production and secretion of pro-angiogenic factors. (A) Representative images of Mouse Angiogenesis Protein Array (R&D) membranes. Squares highlight the spots corresponding to Timp-1 (1), Serpin-E1 (2) and Cxcl-12 (3). (B) Protein quantification of Timp-1, Serpin-E1, and Cxcl-12 in macrophage culture supernatants from M2-polarized MK2-WT or KO macrophages. Spot intensity was quantified in ImageJ. Data corresponds to three membranes per genotype, where culture supernatants from three independent biological replicates were tested. (C) RNA quantification of Timp-1, Serpin-E1, and Cxcl-12 from M2-polarized MK2-WT or KO macrophages from the experiments shown Figure 1 . Box and whisker plots display median (line), 25 th to 75 th percentiles (boxes) and min and maximum (whiskers) RNA expression levels from 6 biological replicates each of MK2-WT and KO macrophages. (D) Representative pictures of angiogenic factors detected by immunohistochemistry in MK2 WT (upper panel, 10 mice) and MK2 KO (lower panel, 5 mice) colon tumors. (E) Stained slides from serial sections of Swiss-rolled entire colons were scanned and quantified in an automated manner using the positive nuclei algorithm in ImageScope and normalized by area analyzed in mm 2 . Each data point corresponds to a single tumor area, and all tumors from all mice with the same genotype [MK2 WT (10 mice) and MK2 KO (5 mice)] are shown. Scale bar 300 um. In panels (B, C, E) statistical significance was

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Caveolin-1 and TIMP-1 are upregulated in cirrhosis and HCC. (A) Representative epifluorescent images of human tissue caveolin-1 and TIMP-1 immunostaining of healthy, cirrhotic, and HCC (hepatocellular carcinoma) liver; protein of interest (red), alpha smooth muscle actin (green) and nuclei (blue); scale bar is 20 mum. Quantification of TIMP-1 (B), and caveolin-1 (C) immunofluorescence staining for panel (A) ( n = number of patients: 5 healthy, 21 cirrhosis, 19 HCC stage I and II, 26 HCC stage III A and B, 7 HCC stage III C and IV A). Whiskers on box and whiskers graphs represent minimum and maximum. Markers denote significant difference from healthy condition by one-way ANOVA with Dunnett's post hoc test, n.s. : not significant, ** 0.001 < p < 0.01, *** 0.0001 < p < 0.001, **** p < 0.0001.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 6 3D self-assembling peptide gels of high fibrosis-mimicking stiffness trigger upregulation of caveolin-1 and TIMP-1. (A) Average storage modulus ( G ') across 0.1-10% strain sweep and (B) elastic modulus of PeptiGels calculated as E = 2 x G ' (1 + upsilon) where, upsilon = Poisson's ratio of 0.5. Histogram bars represent mean +- s.e.m; dots represent individual data points. (C) Representative epifluorescent images of hepatic stellate cells caveolin-1 and TIMP-1 immunostaining of formalin fixed, paraffin embedded soft (4 kPa, gamma 2), and stiff (10 kPa, alpha 2) 3D MBG PeptiGels; protein of interest (red), alpha smooth muscle actin (green) and nuclei (blue), scale bar is 20 mum. TIMP-1 (D) and caveolin-1 (E) immunofluorescence staining for panel (C). TIMP-1 n = 13 and 11, caveolin-1 n = 14 and 18 for gamma 2 and alpha 2 respectively. Mean +- s.e.m. Markers denote significant difference between gamma 2 and alpha 2 by t test, ** 0.001 < p < 0.01, **** p < 0.0001.