Anti-TIMP1 antibody [EPR30559-561]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TIMP1 antibody [EPR30559-561] (AB325798)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling TIMP1 with ab325798 at 1/100 (5.04 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : no staining on mouse cerebrum (PMID : 37095886). The primary antibody was incubated for 30 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TIMP1 antibody [EPR30559-561] (AB325798)

Immunohistochemical analysis of paraffin-embedded Mouse colon of Crohn's disease model tissue labeling TIMP1 with ab325798 at 1/100 (5.04 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse colon of Crohn's disease model. The primary antibody was incubated for 30 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TIMP1 antibody [EPR30559-561] (AB325798)

Immunohistochemical analysis of paraffin-embedded Mouse glioblastoma tissue labeling TIMP1 with ab325798 at 1/100 (5.04 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse glioblastoma. The primary antibody was incubated for 30 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-TIMP1 antibody [EPR30559-561] (AB325798)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized MEF(mouse embryo fibroblast) and Neuro-2a (mouse neuroblastoma neuroblast) cells labelling TIMP1 with ab325798 at 1/50 (10.08 μg/ml) dilution , followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing cytoplasmic staining in MEF cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).

Negative control : Neuro-2a.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 (1 ug/ml) dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 (2 ug/ml) dilution (Magenta).

-ve control 1 : ab325798 at 1/50 dilution, followed by ab150120 at 1/1000 dilution.

-ve control 2 : ab7291 at 1/1000 dilution, followed by ab150081 at 1/1000 dilution.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TIMP1 antibody [EPR30559-561] (AB325798)

Immunohistochemical analysis of paraffin-embedded (A) MEF (mouse embryo fibroblast) cell pellet and (B) Neuro-2a (mouse neuroblastoma neuroblast) cell pellet labeling TIMP1 with ab325798 at 1/100 (5.04 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) MEF cell pellet, no staining on (B) Neuron-2a cell pellet. The primary antibody was incubated for 30 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• WB

Lab

Western blot - Anti-TIMP1 antibody [EPR30559-561] (AB325798)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Low expression : Neuro-2a.

The molecular weight observed is consistent with what has been described in the literature (PMID : 19383295).

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) (1 : 10000) (124KDa).

All lanes:

Western blot - Anti-TIMP1 antibody [EPR30559-561] (ab325798) at 1/1000 dilution

Lane 1:

Untreated MEF (mouse embryo fibroblast) whole cell lysate at 80 µg

Lane 2:

MEF treated with 300ng/ml BFA for 18h whole cell lysate at 80 µg

Lane 3:

Untreated Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 80 µg

Lane 4:

Neuro-2a treated with 300ng/ml BFA for 18h whole cell lysate at 80 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 20-25 kDa,124 kDa

false

Exposure time: 10s

• WB

Lab

Western blot - Anti-TIMP1 antibody [EPR30559-561] (AB325798)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Low expression : cerebrum (PMID : 37095886).

The molecular weight observed is consistent with what has been described in the literature (PMID : 19383295).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

Exposure time : Lanes 1-2 : 180 seconds, Lanes 3-4 : 103 seconds

All lanes:

Western blot - Anti-TIMP1 antibody [EPR30559-561] (ab325798) at 1/1000 dilution

Lane 1:

Rat lung culture supernatant at 80 µg

Lane 2:

Rat cerebellum tissue lysate at 80 µg

Lane 3:

Mouse lung tissue lysate at 80 µg

Lane 4:

Mouse cerebellum tissue lysate at 80 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 20-25 kDa,36 kDa

false