Anti-VGF antibody [EPR27029-73]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VGF antibody [EPR27029-73] (AB308287)

Immunohistochemical analysis of paraffin-embedded Human adrenal gland tissue labelling VGF with ab308287 at 1/500 (1.033 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human adrenal medulla. The section was incubated with ab308287 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-VGF antibody [EPR27029-73] (AB308287)

Immunohistochemical analysis of paraffin-embedded Human pituitary glan tissue labelling VGF with ab308287 at 1/500 (1.033 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human pituitary gland. The section was incubated with ab308287 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-VGF antibody [EPR27029-73] (AB308287)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labelling VGF with ab308287 at 1/500 (1.033 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : no staining on human kidney. The section was incubated with ab308287 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-VGF antibody [EPR27029-73] (AB308287)

Immunohistochemical analysis of paraffin-embedded Rat pituitary gland tissue labelling VGF with ab308287 at 1/500 (1.033 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on rat pituitary gland. The section was incubated with ab308287 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-VGF antibody [EPR27029-73] (AB308287)

Immunohistochemical analysis of paraffin-embedded Mouse adrenal gland tissue labelling VGF with ab308287 at 1/500 (1.033 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse adrenal medulla. The section was incubated with ab308287 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-VGF antibody [EPR27029-73] (AB308287)

Immunohistochemical analysis of paraffin-embedded Rat adrenal gland tissue labelling VGF with ab308287 at 1/500 (1.033 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on rat adrenal medulla. The section was incubated with ab308287 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

• mIHC

Lab

Multiplex immunohistochemistry - Anti-VGF antibody [EPR27029-73] (AB308287)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded rat adrenal gland tissue staining Angiotensin II Type 2 Receptor with ab324679 at a 1/100 ( 5.0 μg/ml) dilution, ab308287 anti-VGF used at a 1/500 ( 0.498 μg/ml) dilution and ab324019 anti-CYP21A2 used at a 1/5000 ( 0.1 μg/ml) dilution.

Panel A : merged staining of anti-Angiotensin II Type 2 Receptor (green; Opal™520), anti-VGF (magenta; Opal™570) and anti-CYP21A2 (gray; Opal™690) on rat adrenal gland.

Panel B : anti-Angiotensin II Type 2 Receptor staining membrane of adrenal medulla in rat adrenal gland.

Panel C : anti-VGF staining endocrine cells of adrenal medulla in rat adrenal gland.

Panel D : anti-CYP21A2 staining adrenal cortex in rat adrenal gland.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab324679, ab308287 and ab324019 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

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

• mIHC

Lab

Multiplex immunohistochemistry - Anti-VGF antibody [EPR27029-73] (AB308287)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse adrenal gland tissue staining Angiotensin II Type 2 Receptor with ab324679 at a 1/100 ( 5.0 μg/ml) dilution, ab308287 anti-VGF used at a 1/500 ( 0.498 μg/ml) dilution and ab324019 anti-CYP21A2 used at a 1/5000 ( 0.1 μg/ml) dilution.

Panel A : merged staining of anti-Angiotensin II Type 2 Receptor (green; Opal™520), anti-VGF (magenta; Opal™570) and anti-CYP21A2 (gray; Opal™690) on mouse adrenal gland.

Panel B : anti-Angiotensin II Type 2 Receptor staining membrane of adrenal medulla in mouse adrenal gland.

Panel C : anti-VGF staining endocrine cells of adrenal medulla in mouse adrenal gland.

Panel D : anti-CYP21A2 staining adrenal cortex in mouse adrenal gland.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab324679, ab308287 and ab324019 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

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

• mIHC

Lab

Multiplex immunohistochemistry - Anti-VGF antibody [EPR27029-73] (AB308287)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse adrenal gland staining of ADX with ab321899 at a 1/2000 (0.244 µg/ml) dilution, VGF with ab308287 at 1/500 (0.95 µg/ml) dilution and Neuropeptide Y with ab221145 at 1/4000 ( 0.140 µg/ml) dilution.

Panel A : merged staining of anti-ADX (gray; Opal™690), anti-VGF (green; Opal™520) and anti-Neuropeptide Y (magenta; Opal™570) on mouse adrenal gland.
Panel B : anti-ADX staining adrenal cortex in mouse adrenal gland.
Panel C : anti-VGF staining endocrine cells in mouse adrenal gland medulla.
Panel D : nti-Neuropeptide Y staining chromaffin cells in mouse adrenal gland medulla.
Nuclear DNA was labelled with DAPI (shown in blue). ).

The section was incubated in three rounds of staining : in the order of ab321899, ab308287 and ab221145 for 30 mins at room temperature.. Each round was followed by a separate fluorescent tyramide signal amplification system.

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

• WB

Supplier Data

Western blot - Anti-VGF antibody [EPR27029-73] (AB308287)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. The primary band at 90 kDa is the precursor form of VGF which is consistent with literature descriptions (PMID : 30690892, PMID : 10433233). In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 180 seconds

All lanes:

Western blot - Anti-VGF antibody [EPR27029-73] (ab308287) at 1/1000 dilution

Lane 1:

Untreated U-2 OS (human bone osteosarcoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

U-2 OS treated with /ml BFA for 24 hour 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

Predicted band size: 67 kDa

Observed band size: 90 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-VGF antibody [EPR27029-73] (AB308287)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. The primary band at 90 kDa is the precursor form of VGF which is consistent with literature descriptions (PMID : 30690892, PMID : 10433233). In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 180 seconds

All lanes:

Western blot - Anti-VGF antibody [EPR27029-73] (ab308287) at 1/1000 dilution

Lane 1:

Untreated PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg

Lane 2:

PC-12 treated with /ml BFA for 24 hour 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

Predicted band size: 67 kDa

Observed band size: 90 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-VGF antibody [EPR27029-73] (AB308287)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. Negative control : RAW 264.7. The identities of the lower MW bands between 37 kDa and 90 kDa are expected to be VGF peptides. The primary band at 90 kDa is the precursor form of VGF which is consistent with literature descriptions (PMID : 30690892, PMID : 10433233). Lysates were freshly made and used for Western blotting immediately to minimize protein degradation. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 180 seconds

All lanes:

Western blot - Anti-VGF antibody [EPR27029-73] (ab308287) at 1/1000 dilution

Lane 1:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 2:

RAW264.7 (mouse elson murine leukemia virus-induced tumor macrophage) 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 1/20000 dilution

Predicted band size: 67 kDa

Observed band size: 90 kDa

false

Exposure time: 180s