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

Rabbit Recombinant Monoclonal VGF antibody. Suitable for WB, IHC-P, ICC/IF, Flow Cyt (Intra), mIHC and reacts with Human, Mouse, Rat samples.

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Images

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

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	WB	IHC-P	ICC/IF	Flow Cyt (Intra)	IP	mIHC
Human	Tested	Tested	Tested	Tested	Not recommended	Expected
Mouse	Tested	Tested	Tested	Tested	Not recommended	Tested
Rat	Tested	Tested	Tested	Not recommended	Not recommended	Tested

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/1000	Notes -
Species Mouse	Dilution info 1/1000	Notes -
Species Rat	Dilution info 1/1000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/500	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Mouse	Dilution info 1/500	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/500	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/50	Notes -
Species Mouse	Dilution info 1/50	Notes -
Species Rat	Dilution info 1/50	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/500	Notes -
Species Mouse	Dilution info 1/500	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human, Mouse, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/500	Notes -
Species Rat	Dilution info 1/500	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Human	Dilution info Use at an assay dependent concentration.	Notes -

Associated Products

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1 product for Alternative Version

Target data

See full target information Neurosecretory protein VGF

Function

Neurosecretory protein VGF. Secreted polyprotein that is packaged and proteolytically processed by prohormone convertases PCSK1 and PCSK2 in a cell-type-specific manner (By similarity). VGF and peptides derived from its processing play many roles in neurogenesis and neuroplasticity associated with learning, memory, depression and chronic pain (By similarity). Neuroendocrine regulatory peptide-1. Plays a role in the control of body fluid homeostasis by regulating vasopressin release. Suppresses presynaptic glutamatergic neurons connected to vasopressin neurons. Neuroendocrine regulatory peptide-2. Plays a role in the control of body fluid homeostasis by regulating vasopressin release. Activates GABAergic interneurons which are inhibitory neurons of the nervous system and thereby suppresses presynaptic glutamatergic neurons (By similarity). Stimulates also feeding behavior in an orexin-dependent manner in the hypothalamus (By similarity). Functions as a positive regulator for the activation of orexin neurons resulting in elevated gastric acid secretion and gastric emptying (By similarity). VGF-derived peptide TLQP-21. Secreted multifunctional neuropeptide that binds to different cell receptors and thereby plays multiple physiological roles including modulation of energy expenditure, pain, response to stress, gastric regulation, glucose homeostasis as well as lipolysis (By similarity). Activates the G-protein-coupled receptor C3AR1 via a folding-upon-binding mechanism leading to enhanced lipolysis in adipocytes (By similarity). Interacts with C1QBP receptor in macrophages and microglia causing increased levels of intracellular calcium and hypersensitivity (By similarity). VGF-derived peptide TLQP-62. Plays a role in the regulation of memory formation and depression-related behaviors potentially by influencing synaptic plasticity and neurogenesis. Induces acute and transient activation of the NTRK2/TRKB receptor and subsequent CREB phosphorylation (By similarity). Induces also insulin secretion in insulinoma cells by increasing intracellular calcium mobilization (By similarity). Antimicrobial peptide VGF[554-577]. Has bactericidal activity against M. luteus, and antifungal activity against P. Pastoris.

Alternative names

Neurosecretory protein VGF, VGF

Recommended products

Rabbit Recombinant Monoclonal VGF antibody. Suitable for WB, IHC-P, ICC/IF, Flow Cyt (Intra), mIHC and reacts with Human, Mouse, Rat samples.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR27029-60
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

VGF also known as non-acronymic VGF is a neuropeptide precursor protein with a molecular mass approximately 68 kDa. It is expressed in neuroendocrine and neuronal tissues including the brain spinal cord and peripheral nervous system. VGF is synthesized as a large precursor and subsequently processed into smaller bioactive peptides with a variety of functions. The distribution and processing of VGF peptides suggest its diverse roles across different physiological contexts.

Biological function summary

VGF-derived peptides exert significant influence on energy balance neurogenesis and synaptic plasticity. These peptides participate in the regulation of appetite and energy expenditure contributing to body weight management. They are also involved in nerve growth and repair mechanisms. VGF peptides act independently or in conjunction with other neuropeptides and factors within the nervous system though it does not form a stable complex with other proteins.

Pathways

Few proteins have as diverse involvement as VGF in the neuroendocrine system's regulation. VGF links to the neurotrophic signaling pathway which promotes neuronal survival and growth. It shows interactions with proteins such as Brain-Derived Neurotrophic Factor (BDNF) and Nerve Growth Factor (NGF) which support cellular health and energy homeostasis. VGF's role assists in the modulation of these pathways facilitating communication between energy state signals and neuronal networks.

Associated diseases and disorders

Alterations in VGF expression have implications in neurodegenerative diseases and mood disorders. Reduced levels of VGF peptides associate with Alzheimer's disease affecting cognitive functions and synaptic integrity. Additionally VGF influences in mood regulatory pathways relate it to major depressive disorder potentially interacting with proteins like the serotonin transporter which modulates mood and stress responses. Understanding VGF and its pathways can inform therapeutic strategies for these conditions.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

20 product images

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 Anti-FSH beta antibody [EPR24439-141] 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 Anti-FSH beta antibody [EPR24439-141] 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.
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 Anti-CYP11A1 antibody [EPR24868-86] ab272494 at a 1/10000 (0.053 µg/ml) dilution and Neuropeptide Y with Anti-Neuropeptide Y antibody [EPR21877] 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, Anti-CYP11A1 antibody [EPR24868-86] ab272494 and Anti-Neuropeptide Y antibody [EPR21877] 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.
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 Anti-CYP11A1 antibody [EPR24868-86] ab272494 at a 1/10000 (0.053 µg/ml) dilution and Neuropeptide Y with Anti-Neuropeptide Y antibody [EPR21877] 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, Anti-CYP11A1 antibody [EPR24868-86] ab272494 and Anti-Neuropeptide Y antibody [EPR21877] 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.
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.
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 (Anti-GAPDH antibody [EPR16891] - Loading Control 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) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 80 kDa, 90 kDa
Exposure time: 180s
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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa FluorÂ® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody. Negative control: RAW 264.7.
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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa FluorÂ® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.
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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed 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). Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.
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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed 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). Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.
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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed 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). Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.
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.
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.
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.
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.
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.
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.
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 (Anti-GAPDH antibody [EPR16891] - Loading Control 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) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 90 kDa
Exposure time: 26s
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 (Anti-GAPDH antibody [EPR16891] - Loading Control 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) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 90 kDa
Exposure time: 26s
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 (Anti-GAPDH antibody [EPR16891] - Loading Control 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) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 90 kDa
Exposure time: 180s
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 (Anti-GAPDH antibody [EPR16891] - Loading Control 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) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 90 kDa
Exposure time: 62s