Anti-CTGF antibody [RM1199]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-CTGF antibody [RM1199] (AB318148)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized CTGF KO U-2 OS (CTGF knockout human bone osteosarcoma epithelial cell) cells labelling CTGF with ab318148 at 1/500 (1.03 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing cytoplasmic staining in wildtype U-2 OS cells (shown in green) and showing negative staining in CTGF knockout U-2 OS cells. The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-CTGF antibody [RM1199] (AB318148)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) (Right) / MCF7 (human breast adenocarcinoma epithelial cell) (Left) cells labelling CTGF with ab318148 at 1/500 dilution (0.1ug) (Red) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Low expression : MCF-7.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-CTGF antibody [RM1199] (AB318148)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) (Right) / MOLT-4 (human lymphoblastic leukemia T lymphoblast) (Left) cells labelling CTGF with ab318148 at 1/500 dilution (0.1ug) (Red) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Low expression : MOLT-4.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CTGF antibody [RM1199] (AB318148)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) cells labelling CTGF with ab318148 at 1/500 (1.03 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing cytoplasmic staining in HepG2 cells (shown in green) . The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Low expression : MOLT-4.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CTGF antibody [RM1199] (AB318148)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Rat-1 (rat embryonic fibroblast) cells labelling CTGF with ab318148 at 1/500 (1.03 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing cytoplasmic staining in Rat-1 cells (shown in green) after starvation 18 hours, then treatment with TGF beta 1(10 ng/ml) and Heparin sodium salt (50 ug/ml) for 24 hours. The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CTGF antibody [RM1199] (AB318148)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling CTGF with ab318148 at 1/500 (1.03 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing cytoplasmic staining in NIH/3T3 cells (shown in green) after starvation 18 hours, then treatment with TGF beta 1(10 ng/ml) and Heparin sodium salt (50 ug/ml) for 24 hours. The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• WB

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Western blot - Anti-CTGF antibody [RM1199] (AB318148)

Low expression : MCF7 and MOLT-4.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 23139278 and PMID : 15955090).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-CTGF antibody [RM1199] (ab318148) at 1/500 dilution

Lane 1:

Untreated Rat-1 (rat embryonic fibroblast) starved for 18 hours, whole cell lysate at 48 µg

Lane 2:

Rat-1 starved for 18 hours, then treated with 10ng /ml TGF--β1 and 50μg/ml Heparin sodium salt for 24 hours, whole cell lysate at 48 µg

Lane 3:

Untreated NIH/3T3 (mouse embryonic fibroblast) starved for 18 hours, whole cell lysate at 48 µg

Lane 4:

NIH/3T3 starved for 18 hours, then treated with 10ng /ml TGF--β1 and 50μg/ml Heparin sodium salt for 24 hours, whole cell lysate at 48 µg

Lane 5:

HFF-1 (human skin fibroblast) whole cell lysate at 48 µg

Lane 6:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 48 µg

Lane 7:

Huh7 (human hepatocellar carcinoma epithelial cell) whole cell lysate at 48 µg

Lane 8:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 48 µg

Lane 9:

MOLT-4 (human lymphoblastic leukemia T lymphoblast) whole cell lysate at 48 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: 38 kDa,18 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-CTGF antibody [RM1199] (AB318148)

Lysates/proteins at 48 µg per lane.

In Western blot, ab318148 was shown to bind specifically to CTGF. A band was observed at 18-38 kDa in wild-type U-2 OS cell lysates with whereas no signal observed at this size in CTGF knockout cell line.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-CTGF antibody [RM1199] (ab318148) at 1/500 dilution

Lane 1:

Wild-type U-2 OS (human bone osteosarcoma epithelial cell) whole cell lysate at 48 µg

Lane 2:

CTGF knockout U-2 OS whole cell lysate at 48 µg

Lane 3:

HepG2 (human hepatocellar carcinoma epithelial cell) whole cell lysate at 48 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: 38 kDa,18 kDa,36 kDa

false

Exposure time: 180s