Anti-CTGF antibody (ab6992)

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

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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.

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.

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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).

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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.

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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.

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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.

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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).

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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.

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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

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