Anti-Adiponectin 抗体 [19F1]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Adiponectin antibody [19F1] (AB22554)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) of Mouse Adipose tissue staining Adiponectin using ab22554 (PBS in negative control) at 1 : 200 (5μg/ml). ImmunoHistoProbe one step HRP Polymer (ready to use) used. Performed heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0)

• ICC

AbReview23243****

Immunocytochemistry - Anti-Adiponectin antibody [19F1] (AB22554)

ab22554 staining Adiponectin in human adipose stem cells by Immunocytochemistry/ Immunofluorescence. The cells were fixed in paraformaldehyde, permeabilised in 0.1% Triton X and then blocked using 4% serum for 1 hour. Samples were then incubated with primary antibody at 1/500 for 1 hour 30 minutes. The secondary antibody used was a goat IgG conjugated to Alexa Fluor® 488 (green) used at a 1/500 dilution.

Image courtesy of an anonymous Abreview.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Adiponectin antibody [19F1] (AB22554)

Immunohistochemistry was performed on cancer biopsies of deparaffinized Human colon carcinoma tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1 : 100 with a mouse monoclonal antibody recognizing Adiponectin ab22554 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Adiponectin antibody [19F1] (AB22554)

Immunohistochemistry was performed on biopsies of deparaffinized Human skin tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1 : 20 with a mouse monoclonal antibody recognizing Adiponectin ab22554 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Adiponectin antibody [19F1] (AB22554)

ab22554 (4µg/ml) staining adiponectin in human breast, using an automated system (DAKO Autostainer Plus). Using this protocol there is nuclear and cytoplasmic staining.
Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.

• ICC

Lab

Immunocytochemistry - Anti-Adiponectin antibody [19F1] (AB22554)

ab22554 staining Adiponectin in 3T3-L1 cells (ATCC® CL-173 TM). Increased expression of Adiponectin correlates with adipocyte phenotype, as described in literature. Cells, grown to confluency in DMEM with 10% FBS, were differentiated by stimulation for two days with 0.5 mM 3-isobutyl-1-methylxanthi​ne (ab120840), 0.25uM dexamethasone (ab120743) and 1ug/ml insulin (ab123768), followed by two more days with only insulin. Cells were maintained for an additional three days in growth medium alone. Undifferentiated and differentiated adipocytes were fixed with 100% methanol (5min) at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 1h at room temperature. Staining of the treated cells with ab22554 (2.5μg/ml) and ab6046 at 1μg/ml overnight at +4°C, was followed by a further incubation at room temperature for 1h with an AlexaFluor®488 Goat anti-Rabbit secondary (ab150081) at 2 μg/ml (shown in green) and AlexaFluor®594 Goat anti-Mouse secondary (ab150120) at 2 μg/ml (shown in pseudo colour red). Nuclear DNA was labelled in blue with DAPI. Negative controls : 1– Rabbit primary and anti-mouse secondary antibody; 2 – Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.

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

• IHC-P

AbReview20510****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Adiponectin antibody [19F1] (AB22554)

ab22554 staining Adiponectin in Mouse skin tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 10% serum for 1 hour at 20°C; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody (1/100) for 12 hours at 4°C. A Biotin-conjugated Rabbit anti-mouse polyclonal (1/200) was used as the secondary antibody.

This image is courtesy of an anonymous Abreview

• WB

Unknown

Western blot - Anti-Adiponectin antibody [19F1] (AB22554)

A ~26 kDa band corresponding Adiponectin was observed in the cell line tested.

All lanes:

Western blot - Anti-Adiponectin antibody [19F1] (ab22554) at 2 µg/mL

All lanes:

Hep G2 cell line

Secondary

All lanes:

Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP at 1/2500 dilution

Predicted band size: 26 kDa

false