Anti-VAPB antibody [EPR27026-50]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VAPB antibody [EPR27026-50] (AB315013)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling VAPB with ab315013 at 1/2000 (0.257 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cytoplasmic staining on human kidney. The section was incubated with ab315013 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed 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-VAPB antibody [EPR27026-50] (AB315013)

Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling VAPB with ab315013 at 1/2000 (0.257 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cytoplasmic staining on human breast carcinoma. The section was incubated with ab315013 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed 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-VAPB antibody [EPR27026-50] (AB315013)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling VAPB with ab315013 at 1/2000 (0.257 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cytoplasmic staining on human liver. The section was incubated with ab315013 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IP

Supplier Data

Immunoprecipitation - Anti-VAPB antibody [EPR27026-50] (AB315013)

VAPB was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate with ab315013 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab315013 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate

Lane 2 : ab315013 IP in HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate

Lane 3 : Rabbit monoclonal lgG (ab172730) instead of ab315013 in HeLa whole cell lysate

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

The lower band at 11 kDa is likely to be the short isoform.

All lanes:

Immunoprecipitation - Anti-VAPB antibody [EPR27026-50] (ab315013) at 1/30 dilution

All lanes:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate

Secondary

All lanes:

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

false

Exposure time: 6s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VAPB antibody [EPR27026-50] (AB315013)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling VAPB with ab315013 at 1/2000 (0.257 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cytoplasmic staining on mouse kidney. The section was incubated with ab315013 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• WB

Supplier Data

Western blot - Anti-VAPB antibody [EPR27026-50] (AB315013)

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

All lanes:

Western blot - Anti-VAPB antibody [EPR27026-50] (ab315013) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

Caco-2 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 27 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-VAPB antibody [EPR27026-50] (AB315013)

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

In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) staining at 1/100000 dilution.

All lanes:

Western blot - Anti-VAPB antibody [EPR27026-50] (ab315013) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

HeLa transfected with siRNA specifically targeting VAPB whole cell lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 27 kDa,15 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-VAPB antibody [EPR27026-50] (AB315013)

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

Exposure Time : Lane 1-3 : 15 seconds, lane 4-5 : 180 seconds.

All lanes:

Western blot - Anti-VAPB antibody [EPR27026-50] (ab315013) at 1/1000 dilution

Lane 1:

Human testis tissue lysate at 20 µg

Lane 2:

Human liver tissue lysate at 20 µg

Lane 3:

Mouse testis tissue lysate at 20 µg

Lane 4:

Mouse liver tissue lysate at 20 µg

Lane 5:

Mouse colon tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Anti-mouse IgG for IP (HRP) (<a href='/en-us/products/secondary-antibodies/mouse-igg-for-ip-hrp-ab131368'>ab131368</a>) at 1/5000 dilution

Observed band size: 27 kDa

false