Invitrogen

TIMP1 Monoclonal Antibody (102D1)

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TIMP1 Antibody in Immunofluorescence (IF) — Immunofluorescent analysis of TIMP-1 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 1% BSA for 1 hour at room temperature. The cells were labeled with TIMP-1 (102D1) Mouse Monoclonal Antibody (Product # MS608PABX) at 1:250 dilution in0.1% 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 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.

TIMP1 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry analysis of TIMP-1 (102D1) showing staining in the cytoplasm of paraffin-embedded human prostate tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a TIMP-1 Antibody (102D1) Mouse Monoclonal Antibody (Product # MS608PABX) diluted in 3% BSA-PBS at a dilution of 1:100 for 1 hour at 37°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

TIMP1 Antibody in Western Blot (WB) — Western blot analysis was performed on (Fig 1) whole cell extracts (30 µg lysate) of HeLa (Lane 1), HT-29 (Lane 2), SK-OV-3 (Lane 3), MCF7 (Lane 4), PC-3 (Lane 5), K-562 (Lane 6) and HCT 116 (Lane 7). Likewise Western blot analysis was performed on (Fig 2) 10 uL HeLa conditioned media (Lane 1) and 10 uL MDA-MB-231 conditioned media (Lane 2). The blots were probed with Anti-TIMP-1 Receptor Mouse Monoclonal Antibody (Product # MS608PABX, 1:500 dilution) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.4 µg/mL, 1:2500 dilution). A 23 kDa band corresponding to TIMP-1 was observed across the cell lines and conditioned media tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0342BOX), 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 using iBind™ Flex Western Starter Kit (Product # SLF2000S). Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Product Details

Tested Applications

Dilution

Immunocytochemistry (ICC)

1:250

Immunofluorescence (IF)

1:250

Immunohistochemistry (Paraffin) (IHC (P))

1:100

Western Blot (WB)

1:500

Published Applications

Western Blot (WB)

See 7 publications below

Immunohistochemistry (Paraffin) (IHC (P))

See 1 publication below

Immunohistochemistry (IHC)

See 26 publications below

Neutralization (Neu)

See 1 publication below

Product Specifications

Species Reactivity

Human

Published species

Human, Mouse

Host / Isotype

Mouse / IgG1

Class

Monoclonal

Type

Antibody

Clone

102D1

Immunogen

Recombinant human TIMP-1.

Conjugate

Unconjugated

Form

Liquid

Purification

Protein G

Storage buffer

PBS, pH 7.4

Contains

no preservative

Storage conditions

-20°C

RRID

AB_417075

Product Specific Information

MS608PABX targets TIMP-1 in immunohistochemistry and western blot applications and shows reactivity with human samples.

The MS608PABX immunogen is recombinant human TIMP-1.

MS608PABX detects TIMP-1 which has a predicted molecular weight of approximately 21 kDa.

Target Information

TIMP1 (metalloproteinase inhibitor 1) is a metalloproteinase inhibitor that functions by forms one-to-one complexes with target metallproteinases, such as collagenases, and irreversible inactivates them by binding to their catalytic zinc cofactor. TIMP1 acts on MMP1, MMP2, MMP3, MMP7, MMP8, MMP9, MMP10, MMP11, MMP12, MMP14, and MMP16. It also functions as a growth factor that regulates cell differenation, migration, and cell death. TIMP1 activates cellular signaling cascades via CD63 and ITGB1.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Protein Aliases: Collagenase inhibitor; EPA; erythroid potentiating activity; Erythroid-potentiating activity; Fibroblast collagenase inhibitor; Metalloproteinase inhibitor 1; TIMP-1; tissue inhibitor of metalloproteinase 1; Tissue inhibitor of metalloproteinases 1

Gene Aliases: CLGI; EPA; EPO; HCI; TIMP; TIMP-1; TIMP1

UniProt ID: (Human) P01033

Entrez Gene ID: (Human) 7076

Function(s)

cell activation platelet degranulation aging positive regulation of cell proliferation response to hormone negative regulation of endopeptidase activity extracellular matrix disassembly response to cytokine wound healing negative regulation of apoptotic process negative regulation of catalytic activity response to peptide hormone negative regulation of membrane protein ectodomain proteolysis cartilage development negative regulation of trophoblast cell migration regulation of integrin-mediated signaling pathway

Process(es)

protease binding cytokine activity protein binding growth factor activity metalloendopeptidase inhibitor activity metal ion binding

TIMP1 Monoclonal Antibody (102D1)

Product Details

Product Specific Information

Target Information

Bioinformatics

Material safety data sheets (MSDS)

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