Anti-LRFN5 抗体 [EPR24417-5] (BSA and Azide free) (ab302511)

All lanes : Anti-LRFN5 antibody [EPR24417-5] (ab302510) at 1/1000 dilution

Lane 1 : Mouse hippocampus tissue lysate
Lane 2 : Mouse heart tissue lysate
Lane 3 : Rat hippocampus tissue lysate
Lane 4 : Rat heart tissue lysate

Lysates/proteins at 20 µg per lane.

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

Developed using the ECL technique.

Performed under non-reducing conditions.

Predicted band size: 79 kDa
Observed band size: 80-100 kDa why is the actual band size different from the predicted?


Exposure time: 48 seconds

This data was developed using ab302510, the same antibody clone in a different buffer formulation.

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

LRFN5 is glycosylated protein. The molecular weights are consistent with what has been described in the literature (PMID: 16828986, 27152329).

Negative control: heart (PMID: 27152329, 27152329).

Samples are non-boiled as boiling may cause protein aggregates.

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

All lanes : Anti-LRFN5 antibody [EPR24417-5] (ab302510) at 1/1000 dilution

Lane 1 : Rat brain tissue lysate
Lane 2 : Mouse brain tissue lysate
Lane 3 : Mouse spinal cord tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Predicted band size: 79 kDa
Observed band size: 80-100 kDa why is the actual band size different from the predicted?


Exposure time: 180 seconds

This data was developed using ab302510, the same antibody clone in a different buffer formulation.

Blocking / Diluting buffer and concentration: 5% NFDM/TBST

LRFN5 is glycosylated protein. The molecular weights are consistent with what has been described in the literature (PMID: 16828986, 27152329).

Samples are non-boiled as boiling may cause protein aggregates.

This data was developed using ab302510, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling LRFN5 with ab302510 at 1/100 dilution (5.16 μg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection kit). Positive staining on mouse cerebrum (PMID: 27152329). The section was incubated with ab302510 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Primary diluent was used instead of primary antibody, followed by a ready to use secondary antibody LeicaDS9800 (Bond™ Polymer Refine Detection kit).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

This data was developed using ab302510, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded rat cerebrum tissue labeling LRFN5 with ab302510 at 1/100 dilution (5.16 μg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection kit). Positive staining on rat cerebrum. The section was incubated with ab302510 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Primary diluent was used instead of primary antibody, followed by a ready to use secondary antibody LeicaDS9800 (Bond™ Polymer Refine Detection kit).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

This data was developed using ab302510, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded mouse cardiac muscle for labeling LRFN5 with ab302510 at 1/100 dilution (5.16 μg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection kit). Negative control: no staining on mouse cardiac muscle (PMID: 27152329). The section was incubated with ab302510 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Primary diluent was used instead of primary antibody, followed by a ready to use secondary antibody LeicaDS9800 (Bond™ Polymer Refine Detection kit).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

This data was developed using ab302510, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded rat cardiac muscle for labeling LRFN5 with ab302510 at 1/100 dilution (5.16 μg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection kit). Negative control: no staining on rat cardiac muscle. The section was incubated with ab302510 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Primary diluent was used instead of primary antibody, followed by a ready to use secondary antibody LeicaDS9800 (Bond™ Polymer Refine Detection kit).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

This data was developed using ab302510, the same antibody clone in a different buffer formulation.

LRFN5 was immunoprecipitated from 10μg of mouse brain tissue lysate with ab302510 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab302510 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used as secondary antibody at 1/5000 dilution.

Lane 1: Mouse brain tissue lysate 10μg (Input).

Lane 2: ab302510 IP in mouse brain tissue lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab302510 in mouse brain tissue lysate.

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

Exposure time: 67 seconds.

This data was developed using ab302510, the same antibody clone in a different buffer formulation.

LRFN5 was immunoprecipitated from 10μg of rat brain tissue lysate with ab302510 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab302510 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used as secondary antibody at 1/5000 dilution.

Lane 1: Rat brain tissue lysate 10μg (Input).

Lane 2: ab302510 IP in rat brain tissue lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab302510 in rat brain tissue lysate.

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

Exposure time: 67 seconds.