Anti-Robo4 antibody [EPR26282-3] (ab300046)

Rabbit Recombinant Monoclonal ROBO4 antibody. Suitable for WB, IHC-P and reacts with Human, Mouse, Rat samples.

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Images

Western blot - Anti-Robo4 antibody [EPR26282-3] (AB300046), expandable thumbnail

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

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

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Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

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

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 1/100	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Mouse	Dilution info 1/100	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/100	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

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

Not recommended
Not recommended

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

Not recommended
Not recommended

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

Not recommended
Not recommended

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

Target data

See full target information ROBO4

Function

Receptor for Slit proteins, at least for SLIT2, and seems to be involved in angiogenesis and vascular patterning. May mediate the inhibition of primary endothelial cell migration by Slit proteins (By similarity). Involved in the maintenance of endothelial barrier organization and function (PubMed:30455415).

Alternative names

UNQ421/PRO3674, ROBO4, Roundabout homolog 4, Magic roundabout

Recommended products

Rabbit Recombinant Monoclonal ROBO4 antibody. Suitable for WB, IHC-P 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: 59% 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: EPR26282-3
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

Roundabout homolog 4 (Robo4) is a protein weighing approximately 107 kDa. It belongs to the immunoglobulin superfamily and is sometimes referred to as Magic Roundabout. Robo4 primarily gets expressed in endothelial cells playing an important function in vascular biology. It is characterized by having two immunoglobulin-like domains and two fibronectin type III domains making it distinct among the other Robo family members.

Biological function summary

Robo4 stabilizes the vascular network interacting actively with the guidance molecule Slit proteins. It is not part of a larger protein complex but engages directly with these ligands to influence endothelial cell migration and vascular permeability. Its role is significant in maintaining the structural integrity of blood vessels ensuring the appropriate cellular responses important for proper vascular function.

Pathways

Robo4 engages in the Slit-Robo signaling pathway that is essential for vascular development and remodeling. This pathway helps inhibit excessive endothelial migration and angiogenesis. Robo4 interacts with proteins like Slit2 which binds to it as a ligand to mediate these effects. Through this pathway Robo4 performs an important function in processes such as vascular homeostasis and angiogenesis regulation.

Associated diseases and disorders

Robo4 appears to have connections to vascular-related conditions including diabetic retinopathy and age-related macular degeneration. The protein influences pathological angiogenesis which is a hallmark in the progression of these diseases. In these contexts Robo4's interaction with Slit2 proves important as it can restrain abnormal vascular growth offering potential therapeutic angles for treatment approaches focused on targeting the Slit-Robo pathway.

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.

12 product images

Western blot - Anti-Robo4 antibody [EPR26282-3] (ab300046)
Blocking / Diluting buffer and concentration:
5% NFDM/TBST
All lanes: Western blot - Anti-Robo4 antibody [EPR26282-3] (ab300046) at 1/1000 dilution
Lane 1: HUVEC (human umbilical vein endothelial cell) whole cell lysate at 20 µg
Lane 2: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 3: HCT 116 (human colorectal carcinoma epithelial cell) 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 1/20000 dilution
Predicted band size: 107 kDa
Observed band size: 60 kDa
Exposure time: 15s
Western blot - Anti-Robo4 antibody [EPR26282-3] (ab300046)
Blocking / Diluting buffer and concentration:5% NFDM/TBST
The molecular weight observed is consistent with what have been described in literature (PMID: 27882935).

Negative controls: HeLa, HCT 116(PMID: 11944987, PMID: 11076864 ).
All lanes: Western blot - Anti-Robo4 antibody [EPR26282-3] (ab300046) at 1/1000 dilution
Lane 1: Human heart tissue lysate at 20 µg
Lane 2: Mouse spleen tissue lysate at 20 µg
Lane 3: Mouse kidney tissue lysate at 20 µg
Lane 4: Rat spleen tissue lysate at 20 µg
Lane 5: Rat kidney tissue lysate at 20 µg
Secondary
All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051);Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG. Secondary ab dilution 1:2000;1:20,000
Predicted band size: 107 kDa
Observed band size: 60 kDa
Exposure time: 15s
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Robo4 antibody [EPR26282-3] (ab300046)
Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Robo4 with abab300046 at 1/100 (3.77 μg/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Positive staining on endothelial cells in human colon. The section was incubated with ab300046 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Robo4 antibody [EPR26282-3] (ab300046)
Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling Robo4 with abab300046 at 1/100 (3.77 μg/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Positive staining on endothelial cells in mouse colon. The section was incubated with ab300046 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Robo4 antibody [EPR26282-3] (ab300046)
Immunohistochemical analysis of paraffin-embedded Mouse pancreatic can tissue labeling Robo4 with abab300046 at 1/100 (3.77 μg/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Positive staining on endothelial cells in mouse pancreatic cancer. The section was incubated with ab300046 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Robo4 antibody [EPR26282-3] (ab300046)
Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling Robo4 with abab300046 at 1/100 (3.77 μg/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection) was used. Positive staining on endothelial cells in rat cerebrum. The section was incubated with ab300046 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Western blot - Anti-Robo4 antibody [EPR26282-3] (ab300046)
Blocking / Diluting buffer and concentration:

5% NFDM/TBST

Negative controls: HeLa, HCT 116 (PMID: 11944987, PMID: 11076864).
All lanes: Western blot - Anti-Robo4 antibody [EPR26282-3] (ab300046) at 1/1000 dilution
Lane 1: HUVEC (human umbilical vein endothelial cell) whole cell lysate at 20 µg
Lane 2: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 3: HCT 116 (human colorectal carcinoma epithelial cell) 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 1/20000 dilution
Predicted band size: 107 kDa
Observed band size: 60 kDa
Exposure time: 15s
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Robo4 antibody [EPR26282-3] (ab300046)
Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling Robo4 with ab300046 at 1/100 (3.77 µg/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection) was used. Positive staining on endothelial cells in rat cerebrum. The section was incubated with ab300046 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Robo4 antibody [EPR26282-3] (ab300046)
Immunohistochemical analysis of paraffin-embedded Mouse pancreatic cancer tissue labeling Robo4 with abab300046 at 1/100 (3.77 µg/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Positive staining on endothelial cells in mouse pancreatic cancer. The section was incubated with ab300046 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Western blot - Anti-Robo4 antibody [EPR26282-3] (ab300046)
Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling Robo4 with ab300046 at 1/100 (3.77 µg/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Positive staining on endothelial cells in mouse colon. The section was incubated with ab300046 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Western blot - Anti-Robo4 antibody [EPR26282-3] (ab300046)
Blocking / Diluting buffer and concentration:

5% NFDM/TBST

The molecular weight observed is consistent with what have been described in literature (PMID: 27882935).

Negative controls: HeLa, HCT 116(PMID: 11944987, PMID: 11076864 ).
All lanes: Western blot - Anti-Robo4 antibody [EPR26282-3] (ab300046) at 1/1000 dilution
Lane 1: Human heart tissue lysate at 20 µg
Lane 2: Mouse spleen tissue lysate at 20 µg
Lane 3: Mouse kidney tissue lysate at 20 µg
Lane 4: Rat spleen tissue lysate at 20 µg
Lane 5: Rat kidney tissue lysate at 20 µg
Secondary
Lanes 1 - 5: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Lanes 1 - 5: Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051);Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 107 kDa
Observed band size: 60 kDa
Exposure time: 15s
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Robo4 antibody [EPR26282-3] (ab300046)
Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Robo4 with abab300046 at 1/100 (3.77 µ/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection) was used. Positive staining on endothelial cells in human colon. The section was incubated with ab300046 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) was used.

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