Anti-Thyroxine Binding Globulin antibody [EPR24453-121]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Thyroxine Binding Globulin antibody [EPR24453-121] (AB280954)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labelling Thyroxine Binding Globulin with ab280954 at 1/300 (5.92 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on the plasma in human colon. The section was incubated with ab280954 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Thyroxine Binding Globulin antibody [EPR24453-121] (AB280954)

Immunohistochemical analysis of paraffin-embedded Human placenta tissue labelling Thyroxine Binding Globulin with ab280954 at 1/300 (5.92 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on the plasma in human placenta. The section was incubated with ab280954 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Thyroxine Binding Globulin antibody [EPR24453-121] (AB280954)

Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HeLa (Human cervix adenocarcinoma epithelial cell, Left) / HepG2 (Human hepatocellular carcinoma epithelial cell, Right) cells labelling Thyroxine Binding Globulin with ab280954 at 1/50 dilution (1ug) (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). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. Negative control : HeLa cells.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Thyroxine Binding Globulin antibody [EPR24453-121] (AB280954)

Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labelling Thyroxine Binding Globulin with ab280954 at 1/300 (5.92 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Negative control : no staining on human skeletal muscle (PMID : 8143919). The section was incubated with ab280954 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Thyroxine Binding Globulin antibody [EPR24453-121] (AB280954)

Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HEK-293T (Human embryonic kidney epithelial cell) cells labelling Thyroxine Binding Globulin with ab280954 at 1/50 dilution (1ug) (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). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. Negative control : HEK-293T cells.

• IP

Supplier Data

Immunoprecipitation - Anti-Thyroxine Binding Globulin antibody [EPR24453-121] (AB280954)

Thyroxine Binding Globulin was immunoprecipitated from 0.35 mg HepG2 (human hepatocellular carcinoma epithelial cell) supernatant with ab280954 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab280954 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HepG2 (human hepatocellular carcinoma epithelial cell) supernatant 10 μl

Lane 2 : ab280954 IP in HepG2 supernatant

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab280954 in HepG2 supernatant

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 3 minutes.

All lanes:

Immunoprecipitation - Anti-Thyroxine Binding Globulin antibody [EPR24453-121] (ab280954)

Predicted band size: 46 kDa

Observed band size: 54 kDa

false

• IP

Supplier Data

Immunoprecipitation - Anti-Thyroxine Binding Globulin antibody [EPR24453-121] (AB280954)

Thyroxine Binding Globulin was immunoprecipitated from 0.35 mg Human lung tissue lysate with ab280954 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab280954 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Human lung tissue lysate 10 ug

Lane 2 : ab280954 IP in Human lung tissue lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab280954 in Human lung tissue lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 3 minutes.

This blot was developed using a higher sensitivity ECL substrate.

All lanes:

Immunoprecipitation - Anti-Thyroxine Binding Globulin antibody [EPR24453-121] (ab280954)

Predicted band size: 46 kDa

Observed band size: 49 kDa,54 kDa

false

• WB

Lab

Western blot - Anti-Thyroxine Binding Globulin antibody [EPR24453-121] (AB280954)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. Samples are non-boiled as boiling may cause protein aggregates. The MW observed is consistent with the literature (PMID : 12107243). Thyroxine-binding globulin is highly expressed in plasma. (PMID : 820495, 8106612). Bands of 54KDa and 49KDa respectively represent the full length and endogenous cleavage form. Exposure time : Lane 1 : 15 seconds Lane 2 : 37 seconds Lane 3, 4 : 109 seconds.

All lanes:

Western blot - Anti-Thyroxine Binding Globulin antibody [EPR24453-121] (ab280954) at 1/1000 dilution

Lane 1:

Human plasma at 20 µL

Lane 2:

Human small intestine tissue lysate at 40 µg

Lane 3:

Human lung tissue lysate 40ug at 40 µg

Lane 4:

Human skeletal muscle lysate at 40 µg

Secondary

Lane 1:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Lanes 2 - 4:

VeriBlot for IP secondary antibody(HRP)(<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Predicted band size: 46 kDa

Observed band size: 49 kDa,54 kDa

false