Anti-TXNIP antibody [EPR14774] - Mouse IgG2b (Chimeric) - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TXNIP antibody [EPR14774] - Mouse IgG2b (Chimeric) - BSA and Azide free (AB232330)

IHC image of TxNIP staining in a section of formalin fixed, paraffin embedded human normal kidney tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab210826, 2μg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab210826).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-TXNIP antibody [EPR14774] - Mouse IgG2b (Chimeric) - BSA and Azide free (AB232330)

ab210826 staining TXNIP in HeLa (Human cervix adenocarcinoma epithelial cell line) by intracellular flow cytometry. Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. The sample was incubated with primary antibody at 1/100 dilution (1μg/ml) (red). An Alexa Fluor^® 488 Goat anti mouse IgG (ab150113) was used at 1/2000 dilution. Rabbit monoclonal IgG (ab172730) was used as isotype control (black). Cell without incubation with primary antibody and secondary antibody (blue).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab210826).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-TXNIP antibody [EPR14774] - Mouse IgG2b (Chimeric) - BSA and Azide free (AB232330)

ab210826 staining TXNIP in 293T (human embryonic kidney epithelial cell line) cells by Immunocytochemistry/Immunofluorescence (ICC/IF). The cells were fixed with 100% Methanol. Samples were incubated with primary antibody at 10µg/ml (1 : 100 dilution). An Alexa Fluor^® 488 Goat Anti-Mouse was used as the secondary antibody at 2µg/ml (ab150113). ab179504, Anti-beta IV Tubulin was used as a counterstain at 2µg/ml and ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) was used as secondary antibody counterstain at 4µg/ml.
For negative control 1, primary antibody was used at a 10µg/ml and ab150080 was used as secondary antibody at 4 µg/ml. For negative control 2, ab179504 was used as a primary antibody at 2µg/ml and ab150113 was used as a secondary antibody at 2 µg/ml. DAPI was used as a nuclear counterstain.   Confocal image showing cytoplasmic staining in 293T cells.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab210826).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-TXNIP antibody [EPR14774] - Mouse IgG2b (Chimeric) - BSA and Azide free (AB232330)

ab210826 stained in HeLa cells. The cells were fixed with 100% methanol (5min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab210826 at 5μg/ml overnight at +4°C. The secondary antibody was ab150177 used at 1 ug/ml for 1hour at room temperature (colored green). ab206369 (Rabbit monoclonal [EPR16774] to beta Tubulin Alexa Fluor^® 594) was used as a counterstaining at a 1/200 dilution for 1hour at room temperature (pseudo-colored red). DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43μM for 1hour at room temperature.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab210826).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-TXNIP antibody [EPR14774] - Mouse IgG2b (Chimeric) - BSA and Azide free (AB232330)

ab210826 stained in NIH3T3 cells. The cells were fixed with 100% methanol (5min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab210826 at 5μg/ml overnight at +4°C. The secondary antibody was ab150177 used at 1 ug/ml for 1hour at room temperature (colored green). ab206369 (Rabbit monoclonal [EPR16774] to beta Tubulin Alexa Fluor^® 594) was used as a counterstaining at a 1/200 dilution for 1hour at room temperature (pseudo-colored red). DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43μM for 1hour at room temperature.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab210826).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TXNIP antibody [EPR14774] - Mouse IgG2b (Chimeric) - BSA and Azide free (AB232330)

IHC image of ab210826 staining in a section of formalin fixed, paraffin embedded mouse normal kidney, using MOM detection kit, ab127055. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins. The section was incubated with ab210826, 2µg/ml, for 15 mins at room temperature.  DAB was used as the chromogen (ab103723), diluted 1/100 and incubated for 10min at room temperature. The section was counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab210826).

• WB

Supplier Data

Western blot - Anti-TXNIP antibody [EPR14774] - Mouse IgG2b (Chimeric) - BSA and Azide free (AB232330)

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-TXNIP antibody [EPR14774] - Mouse IgG2b (Chimeric) - BSA and Azide free (ab232330) at 1/1000 dilution

Lane 1:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 15 µg

Lane 2:

HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates at 15 µg

Lane 3:

NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates at 15 µg

Lane 4:

PC-12 (Rat adrenal gland pheochromocyteoma) whole cell lysates at 15 µg

Secondary

All lanes:

Rabbit polyclonal secondary antibody to mouse IgG – H&L (HRP) at 1/2000 dilution

Predicted band size: 44 kDa

Observed band size: 55 kDa

false

Exposure time: 30s