Anti-Titin antibody [EPR24455-21]

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-Titin antibody [EPR24455-21] (AB284860)

IHC image of Titin staining in a section of human normal frozen heart performed on a Leica Biosystems BOND® RX instrument using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab284860, 0.5ugml, 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. The inset secondary-only control image is taken from an identical assay without primary antibody. 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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Titin antibody [EPR24455-21] (AB284860)

Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labelling Titin with ab284860 at 1/2000 (0.334 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human skeletal muscle. The section was incubated with ab284860 for 10 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Titin antibody [EPR24455-21] (AB284860)

Immunohistochemical analysis of paraffin-embedded Human lung tissue labelling Titin with ab284860 at 1/2000 (0.334 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Negative control : no staining on human lung. The section was incubated with ab284860 for 10 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Titin antibody [EPR24455-21] (AB284860)

Immunohistochemical analysis of paraffin-embedded Human cardiac muscle tissue labelling Titin with ab284860 at 1/2000 (0.334 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human cardiac muscle (PMID : 24725620). The section was incubated with ab284860 for 10 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-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-Titin antibody [EPR24455-21] (AB284860)

Negative control image : IHC image of Titin staining in a section of human normal frozen liver* performed on a Leica Biosystems BOND® RX instrument using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab284860, 0.5ugml, 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. The inset secondary-only control image is taken from an identical assay without primary antibody. *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Titin antibody [EPR24455-21] (AB284860)

Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labelling Titin with ab284860 at 1/4000 (0.167 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse cardiac muscle (PMID : 15752755). The section was incubated with ab284860 for 10 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Titin antibody [EPR24455-21] (AB284860)

Immunohistochemical analysis of paraffin-embedded Rat cardiac muscle tissue labelling Titin with ab284860 at 1/4000 (0.167 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat cardiac muscle. The section was incubated with ab284860 for 10 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

• WB

Lab

Western blot - Anti-Titin antibody [EPR24455-21] (AB284860)

Blocking and diluting buffer and concentration : 5% NFDM/TBSTNegative control : liver, lung (PMID : 15752755)

The multiples bands observed in the WB have been reported in literature (PMID : 25877298)

ab284860 recognizes TTN at a MW >>250 kDa. In skeletal muscle, ab284860 recognizes the complete TTN protein at >>250 kDa as well as the cleavage products of TTN generated by CAPN3, at 13 kDa, 15 kDa, 18 kDa and 45 kDa. In cardiac muscle, ab284860 recognizes the complete TTN protein at >>250 kDa as well as bands at 13 and 150 kDa.

This blot was developed using a higher sensitivity ECL substrate.

Exposure time : 48 seconds

All lanes:

Western blot - Anti-Titin antibody [EPR24455-21] (ab284860) at 1/1000 dilution

Lane 1:

Human heart tissue lysate

Lane 2:

Human skeletal muscle tissue lysate

Lane 3:

Human liver tissue lysate

Lane 4:

Human lu tissue lysate 20

Secondary

All lanes:

VeriBlot for IP secondary antibody(HRP)(<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Predicted band size: 3816 kDa

false

• WB

Lab

Western blot - Anti-Titin antibody [EPR24455-21] (AB284860)

Blocking and diluting buffer and concentration : 5% NFDM/TBSTNegative control : lung (PMID : 15752755)

The multiples bands observed in the WB have been reported in literature (PMID : 25877298)

ab284860 recognizes TTN at a MW >>250 kDa. In skeletal muscle, ab284860 recognizes the complete TTN protein at >>250 kDa as well as the cleavage products of TTN generated by CAPN3, at 13 kDa, 15 kDa, 18 kDa and 45 kDa. In cardiac muscle, ab284860 recognizes the complete TTN protein at >>250 kDa as well as bands at 13 and 150 kDa.

This blot was developed using a higher sensitivity ECL substrate.

Exposure time : 3 minutes

All lanes:

Western blot - Anti-Titin antibody [EPR24455-21] (ab284860) at 1/1000 dilution

Lane 1:

Mouse heart tissue lysate

Lane 2:

Mouse skeletal muscle tissue lysate

Lane 3:

Mouse lu tissue lysate 20

Lane 4:

Rat heart tissue lysate

Lane 5:

Rat skeletal muscle tissue lysate

Lane 6:

Rat lu tissue lysate 20

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 3816 kDa

false