Anti-VGF antibody [EPR27029-60]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VGF antibody [EPR27029-60] (AB313783)

Immunohistochemical analysis of paraffin-embedded human lung tissue labeling VGF with ab313783 at 1/500 (0.96 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : no staining on human lung (PMID : 29209432). The section was incubated with ab313783 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 [EPR27029-60] (AB313783)

Immunohistochemical analysis of paraffin-embedded Human pituitary gland tissue labeling VGF with ab313783 at 1/500 (0.96 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human pituitary gland. The section was incubated with ab313783 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 [EPR27029-60] (AB313783)

Immunohistochemical analysis of paraffin-embedded Human adrenal gland tissue labeling VGF with ab313783 at 1/500 (0.96 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human adrenal medulla. The section was incubated with ab313783 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.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-VGF antibody [EPR27029-60] (AB313783)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized U-2 OS (human bone osteosarcoma epithelial cell) cells labelling VGF with ab313783 at 1/50 (9.6 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing cytoplasmic staining in U-2 OS cells treated with Brefeldin A (BFA, 300 ng/ml) for 24 h, and showing no staining in untreated U-2 OS cells. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5 ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-VGF antibody [EPR27029-60] (AB313783)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized U-2 OS (human bone osteosarcoma epithelial cell) treated with 300ng/ml BFA for 24h (Red) / untreated U-2 OS cells (Green) cells labelling VGF with ab313783 at 1/500 dilution (0.1 ug)/Red and Green (Red) compared with a Rabbit monoclonal IgG (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, ab150081) at 1/5000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VGF antibody [EPR27029-60] (AB313783)

Immunohistochemical analysis of paraffin-embedded rat adrenal gland tissue labeling VGF with ab313783 at 1/500 (0.96 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on rat adrenal medulla. The section was incubated with ab313783 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 [EPR27029-60] (AB313783)

Immunohistochemical analysis of paraffin-embedded mouse adrenal gland tissue labeling VGF with ab313783 at 1/500 (0.96 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse adrenal medulla. The section was incubated with ab313783 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.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-VGF antibody [EPR27029-60] (AB313783)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized PC-12 (rat adrenal gland pheochromocytoma cell) cells labelling VGF with ab313783 at 1/50 (9.6 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing cytoplasmic staining in PC-12 cell line. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5 ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-VGF antibody [EPR27029-60] (AB313783)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Neuro-2a (mouse neuroblastoma neuroblast) cells labelling VGF with ab313783 at 1/50 (9.6 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing cytoplasmic staining in Neuro-2a cell line. Negative control : RAW 264.7. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5 ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-VGF antibody [EPR27029-60] (AB313783)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse adrenal gland staining of VGF with ab313783 at a 1/500 (0.96 µg/ml) dilution, CYP11A1 with ab272494 at a 1/10000 (0.053 µg/ml) dilution and Neuropeptide Y with ab221145 at a 1/4000 (0.140 µg/ml) dilution followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-VGF (green; Opal™690), anti-CYP11A1 (magenta; Opal™520) and anti-Neuropeptide Y (gray; Opal™570) on mouse adrenal gland.
Panel B : anti-VGF staining endocrine cells in mouse adrenal gland medulla.
Panel C : anti-CYP11A1 staining adrenal cortex in mouse adrenal gland.
Panel D : anti-Neuropeptide Y staining endocrine cells in mouse adrenal gland medulla.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab313783, ab272494 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.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-VGF antibody [EPR27029-60] (AB313783)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Rat adrenal gland staining of VGF with ab313783 at a 1/500 (0.96 µg/ml) dilution, CYP11A1 with ab272494 at a 1/10000 (0.053 µg/ml) dilution and Neuropeptide Y with ab221145 at a 1/4000 (0.140 µg/ml) dilution followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-VGF (green; Opal™690), anti-CYP11A1 (magenta; Opal™520) and anti-Neuropeptide Y (gray; Opal™570) on rat adrenal gland.
Panel B : anti-VGF staining endocrine cells in rat adrenal gland medulla.
Panel C : anti-CYP11A1 staining adrenal cortex in rat adrenal gland.
Panel D : anti-Neuropeptide Y staining chromaffin cellsa in rat adrenal gland medulla.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab313783, ab272494 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.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-VGF antibody [EPR27029-60] (AB313783)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage, Left) / Neuro-2a (mouse neuroblastoma neuroblast, Right) cells labelling VGF with ab313783 at 1/500 dilution (0.1 ug) (Red) compared with a Rabbit monoclonal IgG (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, ab150081) at 1/5000 dilution was used as the secondary antibody. Negative control : RAW 264.7.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VGF antibody [EPR27029-60] (AB313783)

Immunohistochemical analysis of paraffin-embedded mouse pituitary gland tissue labeling VGF with ab313783 at 1/500 (0.96 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse pituitary gland. The section was incubated with ab313783 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 [EPR27029-60] (AB313783)

Immunohistochemical analysis of paraffin-embedded rat pituitary gland tissue labeling VGF with ab313783 at 1/500 (0.96 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on rat pituitary gland. The section was incubated with ab313783 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 [EPR27029-60] (AB313783)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse pituitary gland staining of VGF with ab313783 at a 1/500 (0.96 µg/ml) dilution and FSH-beta with ab281562 at a 1/4000 (0.126 µg/ml) dilution followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-VGF (green; Opal™520), anti-FSH-beta (magenta; Opal™570) on mouse pituitary gland.
Panel B : anti-VGF staining endocrine cells in mouse pituitary gland.
Panel C : anti-FSH-beta staining corticotrophs in mouse pituitary gland.
Panel D : Nuclear DNA was labelled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab313783 and ab281562 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 [EPR27029-60] (AB313783)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : Jurkat (PMID : 24277232). 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 [EPR27029-60] (ab313783) at 1/1000 dilution

Lane 1:

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

Lane 2:

Jurkat (human T cell leukemia T lymphocyte) 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: 90 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-VGF antibody [EPR27029-60] (AB313783)

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. The identity of the band at 20 kDa is unknown. Exposure time : 62 seconds

All lanes:

Western blot - Anti-VGF antibody [EPR27029-60] (ab313783) 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

Observed band size: 90 kDa

false

Exposure time: 62s

• WB

Supplier Data

Western blot - Anti-VGF antibody [EPR27029-60] (AB313783)

Blocking and diluting buffer and concentration : 5% NFDM/TBST 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). In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 26 seconds

All lanes:

Western blot - Anti-VGF antibody [EPR27029-60] (ab313783) at 1/1000 dilution

All lanes:

PC-12 (rat adrenal gland pheochromocytoma) 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: 90 kDa

false

Exposure time: 26s

• WB

Supplier Data

Western blot - Anti-VGF antibody [EPR27029-60] (AB313783)

Blocking and diluting buffer and concentration : 5% NFDM/TBST The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 30690892). Low expression : kidney. 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 [EPR27029-60] (ab313783) at 1/1000 dilution

Lanes 1 and 5:

Mouse adrenal gland tissue lysate at 20 µg

Lane 2:

Mouse kidney tissue lysate at 20 µg

Lanes 3 and 6:

Rat adrenal gland tissue lysate at 20 µg

Lane 4:

Rat kidney tissue 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: 80 kDa,90 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-VGF antibody [EPR27029-60] (AB313783)

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). In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 26 seconds

All lanes:

Western blot - Anti-VGF antibody [EPR27029-60] (ab313783) at 1/1000 dilution

Lane 1:

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

Lane 2:

RAW264.7 (mouse Abelson 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 1/100000 dilution

Observed band size: 90 kDa

false

Exposure time: 26s