Anti-TIMP1 antibody [EPR18352] - BSA and Azide free

• WB

Lab

Western blot - Anti-TIMP1 antibody [EPR18352] - BSA and Azide free (AB219471)

This data was developed using the same antibody clone in a different buffer formulation (ab211926). False colour image of Western blot : Anti-TIMP1 antibody [EPR18352] staining at 1/1000 dilution, shown in black. In Western blot, ab211926 was shown to bind specifically to TIMP1. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times before development with Optiblot (ECL reagent ab133456) and imaged with 20 minutes exposure time. Secondary antibodies used were HRP conjugated Goat anti-Rabbit (H+L) at 1/50000 dilution and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-TIMP1 antibody [EPR18352] (<a href='/en-us/products/primary-antibodies/timp1-antibody-epr18352-ab211926'>ab211926</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lanes 1 - 5:

Western blot - Human TIMP1 knockout HeLa cell line (ab264022)

Lane 2:

TIMP1 knockout HeLa cell lysate at 20 µg

Lane 3:

MOLT-4 cell lysate at 20 µg

Lane 4:

SH-SY5Y cell lysate at 20 µg

Lane 5:

U-87 MG cell lysate at 20 µg

Secondary

Lanes 1 - 5:

Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

Lanes 1 - 5:

Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Predicted band size: 23 kDa

Observed band size: 30 kDa

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• WB

Lab

Western blot - Anti-TIMP1 antibody [EPR18352] - BSA and Azide free (AB219471)

This data was developed using the same antibody clone in a different buffer formulation (ab211926).

Lanes 1-4 : Merged signal (red and green). Green - ab211926 observed at 26 kDa. Red - loading control ab7291 observed at 50 kDa.

ab211926 Anti-TIMP1 antibody [EPR18352] was shown to specifically react with TIMP1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261740 (knockout cell lysate ab257291) was used. Wild-type and TIMP1 knockout samples were subjected to SDS-PAGE. ab211926 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-TIMP1 antibody [EPR18352] (<a href='/en-us/products/primary-antibodies/timp1-antibody-epr18352-ab211926'>ab211926</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

TIMP1 knockout HeLa cell lysate at 20 µg

Lane 3:

HT-1080 treated with 200ng/ml 12-O-Tetradecanoylphorbol-13-acetate (TPA) for 24 hours, cell lysate at 20 µg

Lane 4:

Untreated HT-1080 cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 23 kDa

Observed band size: 26 kDa

false

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-TIMP1 antibody [EPR18352] - BSA and Azide free (AB219471)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HT-29 (Human colorectal adenocarcinoma cell line) cells labeling TIMP1 with ab211926 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HT-29 cell line. The nuclear counter stain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/1000 dilution (red).

The negative controls are as follows :
-ve control 1 : ab211926 at 1/500 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/1000 dilution.
-ve control 2 : Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab211926).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TIMP1 antibody [EPR18352] - BSA and Azide free (AB219471)

Immunohistochemical analysis of paraffin-embedded human lung cancer tissue labeling TIMP1 with ab211926 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on human lung cancer is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab211926).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-TIMP1 antibody [EPR18352] - BSA and Azide free (AB219471)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SK-OV-3 (Human ovarian cancer cell line) cells labeling TIMP1 with ab211926 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on SK-OV-3 cell line. The nuclear counter stain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/1000 dilution (red).

The negative controls are as follows :
-ve control 1 : ab211926 at 1/500 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/1000 dilution.
-ve control 2 : Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab211926).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TIMP1 antibody [EPR18352] - BSA and Azide free (AB219471)

Immunohistochemical analysis of paraffin-embedded human pancreas tissue labeling TIMP1 with ab211926 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on human islet is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab211926).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TIMP1 antibody [EPR18352] - BSA and Azide free (AB219471)

Immunohistochemical analysis of paraffin-embedded human prostate cancer tissue labeling TIMP1 with ab211926 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on human prostate cancer is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab211926).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TIMP1 antibody [EPR18352] - BSA and Azide free (AB219471)

Immunohistochemical analysis of paraffin-embedded human medullary thyroid carcinoma tissue labeling TIMP1 with ab211926 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on human medullary thyroid carcinoma is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab211926).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TIMP1 antibody [EPR18352] - BSA and Azide free (AB219471)

Immunohistochemical analysis of paraffin-embedded human colon tissue labeling TIMP1 with ab211926 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on human colon neuroendocrine cell is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab211926).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.