AB326048

Anti-Robo1 antibody [EPR29534-534]

RabMAb
Recombinant
20ul selling size
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Rabbit Recombinant Monoclonal ROBO1 antibody. Suitable for IHC-P, WB and reacts with Transfected cell line - Human, Human, Mouse, Rat samples.

View Alternative Names

DUTT1, ROBO1, Roundabout homolog 1, Deleted in U twenty twenty, H-Robo-1

14 Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Robo1 antibody [EPR29534-534] (AB326048)

IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Robo1 antibody [EPR29534-534] (AB326048)

Immunohistochemical analysis of paraffin-embedded Human liver cancer tissue labeling Robo1 with ab326048 at 1/100 (5.09 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human liver cancer (PMID : 28977866). The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Robo1 antibody [EPR29534-534] (AB326048)

Immunohistochemical analysis of paraffin-embedded Human adult brain tissue labeling Robo1 with ab326048 at 1/100 (5.09 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : weak staining on human adult brain. The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Robo1 antibody [EPR29534-534] (AB326048)

Immunohistochemical analysis of paraffin-embedded (A) HeLa (human cervical adenocarcinoma epithelial cell) cell pellet (B) U-87 MG (human glioblastoma-astrocytoma epithelial cell) cell pellet (C) HAP1 (human chronic myelogenous leukemia near-haploid cell) cell pellet (D) SW480 (human colorectal adenocarcinoma epithelial cell) cell pellet (E) MCF7 (human breast adenocarcinoma epithelial cell) cell pellet (F) LoVo (human colorectal adenocarcinoma epithelial cell) cell pellet tissue labeling Robo1 with ab326048 at 1/100 (5.09 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) HeLa cell pellet, (B) U-87 MG cell pellet, (C) HAP1 cell pellet, (D) SW480 cell pellet, and a few cells of (E) MCF7 cell pellet, no staining on (F) LoVo cell pellet. The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Robo1 antibody [EPR29534-534] (AB326048)

Immunohistochemical analysis of paraffin-embedded Mouse adult brain tissue labeling Robo1 with ab326048 at 1/100 (5.09 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : weak staining on mouse adult brain. The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Robo1 antibody [EPR29534-534] (AB326048)

Immunohistochemical analysis of paraffin-embedded Mouse neonatal (postnatal day 0) brain tissue labeling Robo1 with ab326048 at 1/100 (5.09 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse neonatal (postnatal day 0) brain (PMID : 12123796). The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Robo1 antibody [EPR29534-534] (AB326048)

Immunohistochemical analysis of paraffin-embedded Rat adult brain tissue labeling Robo1 with ab326048 at 1/100 (5.09 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : weak staining on rat adult brain. The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Robo1 antibody [EPR29534-534] (AB326048)

Immunohistochemical analysis of paraffin-embedded Rat neonatal (postnatal day 0) brain tissue labeling Robo1 with ab326048 at 1/100 (5.09 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat neonatal (postnatal day 0) brain. The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Robo1 antibody [EPR29534-534] (AB326048)

Immunohistochemical analysis of paraffin-embedded Mouse neonatal (postnatal day 5) brain tissue labeling Robo1 with ab326048 at 1/100 (5.09 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse neonatal (postnatal day 5) brain. The primary antibody was incubated 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

Lab

Western blot - Anti-Robo1 antibody [EPR29534-534] (AB326048)

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

Low expression : MCF7, LoVo.

250 kDa full length band and 120-130 kDa cleavage fragment band were ovserved (PMID : 20471383).

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) (1 : 10000) (124KDa).

All lanes:

Western blot - Anti-Robo1 antibody [EPR29534-534] (ab326048) at 1/1000 dilution

Lane 1:

HAP1 (human chronic myelogenous leukemia near-haploid cell) whole cell lysate at 20 µg

Lane 2:

U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

LoVo (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 6:

SW480 (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 1/20000 dilution

Observed band size: 120-130 kDa,250 kDa,124 kDa

false

Exposure time: 37s

Lab

Western blot - Anti-Robo1 antibody [EPR29534-534] (AB326048)

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

Low expression : spleen.

250 kDa full length band and 120-130 kDa cleavage fragment band were ovserved (PMID : 20471383).

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) (1 : 10000) (124KDa).

All lanes:

Western blot - Anti-Robo1 antibody [EPR29534-534] (ab326048) at 1/1000 dilution

Lane 1:

Human breast cancer tissue lysate at 20 µg

Lane 2:

Human liver cancer tissue lysate at 20 µg

Lane 3:

Human colon cancer tissue lysate at 20 µg

Lane 4:

Human spleen tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 120-130 kDa,124 kDa

false

Exposure time: 37s

Lab

Western blot - Anti-Robo1 antibody [EPR29534-534] (AB326048)

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

Low expression : spleen, brain(PMID : 12123796, PMID : 9608531).

250 kDa full length band and 120-130 kDa cleavage fragment band were ovserved (PMID : 20471383).

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) (1 : 10000) (124KDa).

All lanes:

Western blot - Anti-Robo1 antibody [EPR29534-534] (ab326048) at 1/1000 dilution

Lane 1:

Rat P14 brain tissue lysate at 20 µg

Lane 2:

Rat P14 spinal cord tissue lysate at 20 µg

Lane 3:

Rat brain tissue lysate at 20 µg

Lane 4:

Rat spleen tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 120-130 kDa,250 kDa,124 kDa

false

Exposure time: 10s

Lab

Western blot - Anti-Robo1 antibody [EPR29534-534] (AB326048)

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

Low expression : spleen, brain(PMID : 12123796, PMID : 9608531).

250 kDa full length band and 120-130 kDa cleavage fragment band were ovserved (PMID : 20471383).

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) (1 : 10000) (124KDa).

All lanes:

Western blot - Anti-Robo1 antibody [EPR29534-534] (ab326048) at 1/1000 dilution

Lane 1:

Mouse P0 brain tissue lysate at 20 µg

Lane 2:

Mouse P5 brain tissue lysate at 20 µg

Lane 3:

Mouse brain tissue lysate at 20 µg

Lane 4:

Mouse spleen tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 120-130 kDa,250 kDa,124 kDa

false

Exposure time: 10s

IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Robo1 antibody [EPR29534-534] (AB326048)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human embryonic kidney epithelial cell) transfected with a human ROBO1 expression vector containing a his tag (B) HEK-293T transfected with a human ROBO2 expression vector containing a his tag (C) HEK-293T transfected with a human ROBO3 expression vector containing a his tag (D) HEK-293T transfected with a human ROBO4 expression vector containing a his tag (E) HEK-293T transfected with empty vector containing a his tag tissue labeling Robo1 with ab326048 at 1/2000 (0.255 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) HEK-293T transfected with a human ROBO1 expression vector containing a his tag, almost no staining on (B) HEK-293T transfected with a human ROBO2 expression vector containing a his tag, (C) HEK-293T transfected with a human ROBO3 expression vector containing a his tag, (D) HEK-293T transfected with a human ROBO4 expression vector containing a his tag, and (E) HEK-293T transfected with empty vector containing a his tag. The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Robo1 antibody [EPR29534-534] (AB326048)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling Robo1 with ab326048 at 1/100 (5.09 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on mouse spleen. The primary antibody was incubated 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

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR29534-534

Isotype

IgG

Carrier free

Reacts with

Human, Mouse, Rat

Applications

WB, IHC-P

applications

Immunogen

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

Specificity

This antibody exhibits a relatively low tumor positivity rate in IHC‑P.

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "IHCFr" : {"fullname" : "Immunohistochemistry (Frozen sections)", "shortname":"IHC-Fr"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "IHCP-species-checked": "testedAndGuaranteed", "IHCP-species-dilution-info": "1/100", "IHCP-species-notes": " Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "", "IHCFr-species-checked": "notRecommended", "IHCFr-species-dilution-info": "", "IHCFr-species-notes": "" }, "Mouse": { "IHCP-species-checked": "testedAndGuaranteed", "IHCP-species-dilution-info": "1/100", "IHCP-species-notes": " Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "", "IHCFr-species-checked": "notRecommended", "IHCFr-species-dilution-info": "", "IHCFr-species-notes": "" }, "Rat": { "IHCP-species-checked": "testedAndGuaranteed", "IHCP-species-dilution-info": "1/100", "IHCP-species-notes": " Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "", "IHCFr-species-checked": "notRecommended", "IHCFr-species-dilution-info": "", "IHCFr-species-notes": "" }, "Transfected cell line - Human": { "IHCP-species-checked": "testedAndGuaranteed", "IHCP-species-dilution-info": "1/2000", "IHCP-species-notes": " Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "IHCFr-species-checked": "notRecommended", "IHCFr-species-dilution-info": "", "IHCFr-species-notes": "" } } }

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Product details

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

What are the advantages of a recombinant monoclonal antibody?
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.

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Properties and storage information

Form

Liquid

Purification technique

Affinity purification Protein A

Storage buffer

pH: 7.2 - 7.4 Preservative: 0.01% Sodium azide Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

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

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:

Visit the General protocols
Visit the Troubleshooting

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Target data

Receptor for SLIT1 and SLIT2 that mediates cellular responses to molecular guidance cues in cellular migration, including axonal navigation at the ventral midline of the neural tube and projection of axons to different regions during neuronal development (PubMed : 10102268, PubMed : 24560577). Interaction with the intracellular domain of FLRT3 mediates axon attraction towards cells expressing NTN1 (PubMed : 24560577). In axon growth cones, the silencing of the attractive effect of NTN1 by SLIT2 may require the formation of a ROBO1-DCC complex (By similarity). Plays a role in the regulation of cell migration via its interaction with MYO9B; inhibits MYO9B-mediated stimulation of RHOA GTPase activity, and thereby leads to increased levels of active, GTP-bound RHOA (PubMed : 26529257). May be required for lung development (By similarity).

See full target information ROBO1

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com