Anti-CTGF antibody (ab6992)

4% paraformaldehyde-fixed mouse kidney tissue stained for CTGF using ab6992 at 1/250 dilution in immunohistochemical analysis.

The samples were processed for indirect immune detection using a mousse and rabbit specific horseradish peroxidase (HRP)/ diaminobenzidine (DAB) detection IHC kit (ab80436) according to the manufactor’s protocol. Negative controls were included in each staining series, by omission of the primary antibodies (E₁-F₁). Antigen retrieval was performed for 20 min for paraffin-embedded tissue in the preheated Citrate Buffer (10mM Citric Acid, pH 6.0) using a pressure cooker. Between incubations with the antibodies, the specimens were washed two to four times in buffer PBS (pH 7.4). All incubations were performed overnight at 4°C in a humidified chamber. E_1-2  shows CTGF staining in the renal cortex. F_1-2 shows staining in the medulla.

All lanes : Anti-CTGF antibody (ab6992) at 1/1000 dilution

Lane 1 : Extracellular matric within skeletal muscle from a mouse with fibrosis in the legs
Lane 2 : Human skeletal muscle whole tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Alexa Fluor® 488-conjugated goat anti-rabbit IgG at 1/10000 dilution

Performed under reducing conditions.

Observed band size: 36 kDa why is the actual band size different from the predicted?


Exposure time: 2 minutes

See Abreview

ab6992 staining GTGF (green) in Human vasculature tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with mathacarn and blocked with 10% serum for 20 minutes at 25°C. Samples were incubated with primary antibody (1/400) for 14 hours at 4°C. An Alexa Fluor® 488-conjugated Goat anti-rabbit IgG polyclonal (1/400) was used as the secondary antibody. Blue - DAPI Nuclei.

See Abreview

ab6992 at a 1/200 dilution staining CTGF in mouse liver tissue sections by Immunohistochemistry (frozen sections) incubated at +4°C for 9 hours. Fixed in formaldehyde, permeabilized using 0.2% Triton X-100. Blocked using 2% BSA for 30 minutes at 20°C. Secondary used at 1/200 dilution polyclonal Goat anti-rabbit IgG conjugated to Alexa Fluor 488 (green).

See Abreview

ab6992 staining CTGF in Human 293FT cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilzed with 0.1% Triton X-100 and blocked with 5% serum for 1 hour at 25°C. Samples were incubated with primary antibody (1/200 in PBS) for 2 hours at 25°C. An Alexa Fluor^®488-conjugated goat anti-rabbit IgG polyclonal (1/250) was used as the secondary antibody.

See Abreview

ab6992 at a 1/100 dilution staining CTGF from Mouse Tissue sections (Skin) by Immunohistochemistry (Formalin-fixed paraffin-embedded sections). The antibody was incubated with the tissue for 12 hours and then detected using a Labeled AP polymer. The right side of image is regrown wounded epidermis and shows significant staining for CTGF.

See Abreview

ab6992 staining GTGF (green) in Mouse vasculature tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with mathacarn and blocked with 10% serum for 20 minutes at 25°C. Samples were incubated with primary antibody (1/400) for 14 hours at 4°C. An Alexa Fluor® 488-conjugated Goat anti-rabbit IgG polyclonal (1/400) was used as the secondary antibody. Blue - DAPI Nuclei.

See Abreview

ab6692 staining CTGF in rat bone marrow cells by Immunocytochemistry/ Immunofluorescence. The cells were paraformaldehyde fixed, permeabilised in 0.1% Triton X-100 and then blocked using 5% serum for 1 hour at 25°C. Samples were then incubated with primary antibody at 1/200 for 12 hours at 4°C. The secondary antibody used was a goat anti-rabbit IgG conjugated to Alexa Fluor® 488 (green) used at a 1/250 dilution. DAPI was used to stain the cell nuclei (blue).

See Abreview

ab6992 staining CTGF in Sheep articular cartilage tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with 10% neutral-buffered formalin and blocked with 0.25% serum-free protein block for 10 minutes at room temperature. Samples were incubated with primary antibody (1/400) for 16 hours at 4°C. A Biotin-conjugated Goat anti-rabbit IgG polyclonal (1/50) was used as the secondary antibody.

See Abreview

Anti-CTGF antibody (ab6992) at 1/5000 dilution (in PBS-T for 9 hours at 4°C) + Mouse brain whole cell lysate at 50 µg

Secondary
An HRP-conjugated Goat anti-rabbit IgG polyclonal at 1/5000 dilution

Developed using the ECL technique.

Observed band size: 36 kDa why is the actual band size different from the predicted?


Exposure time: 30 seconds


Blocking Step: 5% Milk for 1 hour at 25°C

See Abreview