Anti-Thyroxine Binding Globulin antibody [EPR24453-121]

8 product images

• 

  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 (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 (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.

• 

  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)(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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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

• 

  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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody. Negative control: HeLa cells.

• 

  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)(VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/1000 dilution

  Predicted band size: 46 kDa

  Observed band size: 49 kDa, 54 kDa

• 

  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)(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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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

• 

  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 (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 (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.

• 

  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 (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 (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 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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody. Negative control: HEK-293T cells.