Anti-Rat EphA5 (Clone 86731)

Product No.: E254


Clone 86731 Target EPHA5 Formats AvailableView All Product Type Monoclonal Antibody Alternate Names TYRO4, CEK7, EHK1, HEK7, EPH Receptor A5 Isotype IgG1 Applications CyTOF® , FC , ICC , WB


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Antibody Details Product Details Reactive Species Rat Host Species Mouse Immunogen NS0-Derived Recombinant Rat EphA5 Extracellular Domain Endotoxin Level <0.1 EU/µg as determined by the LAL method Formulation This monoclonal antibody has been 0.2 µm filtered and lyophilized from modified Dulbecco's phosphate buffered saline (1X PBS) pH 7.2 - 7.3 containing 5.0% w/v trehalose with no calcium, magnesium or preservatives present. Storage and Handling The lyophilized antibody can be stored desiccated at -20°C to -70°C for up to twelve months. The reconstituted antibody can be stored for at least four weeks at 2-8°C. For long-term storage of the reconstituted antibody, aseptically aliquot into working volumes and store at -20°C to -70°C in a manual defrost freezer. Avoid repeated freeze thaw cycles. No detectable loss of activity was observed after six months. Country of Origin USA Shipping Next Day Ambient RRIDAB_2830026 Applications and Recommended Usage? Quality Tested by Leinco Flow Cytometry: It is recommended to use the indirect method for signal enhancement when enumerating cells expressing EphA5. A suggested method would be to stain cells expressing EphA5 with 0.25 µg per 1 - 2.5 x 10⁶ cells in a total staining volume of ≤200 µl followed by PN:M1189. Western Blotting: To detect Rat EphA5 this monoclonal antibody can be used at a concentration of 1-2 µg/ml. This monoclonal antibody should be used in conjunction with compatible second-step reagents such as PN:M114 and a chromogenic substrate such as PN:T343. The detection limit for Rat EphA5 is 1 ng/lane under either reducing or non-reducing conditions. The sensitivity of detection may increase up to 50 fold when a chemiluminescent substrate is used. Additional Applications Reported In Literature ? Immunocytochemistry: This antibody was used at a concentration of 10 μg/mL to detect EphA5 on U-118-MG cells. Cells were fixed with PBS containing 4% paraformaldehyde for 20 minutes at room temperature and blocked with PBS containing 10% normal donkey serum, 0.1% Triton X-100, and 1% BSA for 45 minutes at room temperature. After blocking, cells were incubated with diluted primary antibody overnight at 4° C followed by Rhodamine Red™-coupled anti-mouse IgG at room temperature in the dark for one hour. Between each step, cells were washed with PBS containing 0.1% BSA. CyTOF-ready: Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation. Each investigator should determine their own optimal working dilution for specific applications. See directions on lot specific datasheets, as information may periodically change. Description Description Specificity Mouse Anti-Rat EphA5 (Clone 86731) recognizes an epitope on Rat EphA5. This monoclonal antibody was purified using multi-step affinity chromatography methods such as Protein A or G depending on the species and isotype. No cross reactivity was detected with rhEphA1, rmEphA2, -A3, -A6, -A7, rrEphB1, -B2 or rmEphB3. Background EPH receptor A5 (EPHA5) is a glycoprotein that is expressed by astrocytes and skeletal muscle. It provides guidance cues for retinal ganglion cells¹ and is essential for proper retinal axon guidance and topographic mapping in the mammalian visual system.² Ephrin A5 serves as a general inhibitor of axonal growth from limbic neurons.3 It also plays a role in motor axon outgrowth and pathfinding to the hindlimb.^4,5 PubMed EPHA5 NCBI Gene Bank ID 79208 UniProt.org Information on Uniprot.org References & Citations 1. Davenport, RW. et al. (1998) J Neurosci. 18: 975 2. Frisen, J. et al. (1998) Neuron 20: 235 3. Gao, PP. et al. (1998) Proceedings of the National Acad of Sci. 95: 5329 4. Eberhart, J. et al. (2000) Developmental Neurosci. 22: 237 5. Eberhart, J. et al. (2004) J of Neurosci. 24: 1070 View product citations for antibody E254 on CiteAb Technical Protocols CyTOF® ICC Certificate of Analysis Request COA

Antibody Details

Product Details

Reactive Species

Rat

Host Species

Mouse

Immunogen

NS0-Derived Recombinant Rat EphA5 Extracellular Domain

Endotoxin Level

<0.1 EU/µg as determined by the LAL method

Formulation

This monoclonal antibody has been 0.2 µm filtered and lyophilized from modified Dulbecco's phosphate buffered saline (1X PBS) pH 7.2 - 7.3 containing 5.0% w/v trehalose with no calcium, magnesium or preservatives present.

Storage and Handling

The lyophilized antibody can be stored desiccated at -20°C to -70°C for up to twelve months. The reconstituted antibody can be stored for at least four weeks at 2-8°C. For long-term storage of the reconstituted antibody, aseptically aliquot into working volumes and store at -20°C to -70°C in a manual defrost freezer. Avoid repeated freeze thaw cycles. No detectable loss of activity was observed after six months.

Country of Origin

USA

Shipping

Next Day Ambient

RRIDAB_2830026

Applications and Recommended Usage?
Quality Tested by Leinco

Flow Cytometry: It is recommended to use the indirect method for signal enhancement when enumerating cells expressing EphA5. A suggested method would be to stain cells expressing EphA5 with 0.25 µg per 1 - 2.5 x 10⁶ cells in a total staining volume of ≤200 µl followed by PN:M1189.
Western Blotting: To detect Rat EphA5 this monoclonal antibody can be used at a concentration of 1-2 µg/ml. This monoclonal antibody should be used in conjunction with compatible second-step reagents such as PN:M114 and a chromogenic substrate such as PN:T343. The detection limit for Rat EphA5 is 1 ng/lane under either reducing or non-reducing conditions. The sensitivity of detection may increase up to 50 fold when a chemiluminescent substrate is used.

Additional Applications Reported In Literature ?

Immunocytochemistry: This antibody was used at a concentration of 10 μg/mL to detect EphA5 on U-118-MG cells. Cells were fixed with PBS containing 4% paraformaldehyde for 20 minutes at room temperature and blocked with PBS containing 10% normal donkey serum, 0.1% Triton X-100, and 1% BSA for 45 minutes at room temperature. After blocking, cells were incubated with diluted primary antibody overnight at 4° C followed by Rhodamine Red™-coupled anti-mouse IgG at room temperature in the dark for one hour. Between each step, cells were washed with PBS containing 0.1% BSA.
CyTOF-ready: Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Each investigator should determine their own optimal working dilution for specific applications. See directions on lot specific datasheets, as information may periodically change.

Description

Specificity

Mouse Anti-Rat EphA5 (Clone 86731) recognizes an epitope on Rat EphA5. This monoclonal antibody was purified using multi-step affinity chromatography methods such as Protein A or G depending on the species and isotype. No cross reactivity was detected with rhEphA1, rmEphA2, -A3, -A6, -A7, rrEphB1, -B2 or rmEphB3.

Background

EPH receptor A5 (EPHA5) is a glycoprotein that is expressed by astrocytes and skeletal muscle. It provides guidance cues for retinal ganglion cells¹ and is essential for proper retinal axon guidance and topographic mapping in the mammalian visual system.² Ephrin A5 serves as a general inhibitor of axonal growth from limbic neurons.3 It also plays a role in motor axon outgrowth and pathfinding to the hindlimb.^4,5

PubMed

EPHA5

NCBI Gene Bank ID

79208

UniProt.org

Information on Uniprot.org

References & Citations

1. Davenport, RW. et al. (1998) J Neurosci. 18: 975
2. Frisen, J. et al. (1998) Neuron 20: 235
3. Gao, PP. et al. (1998) Proceedings of the National Acad of Sci. 95: 5329
4. Eberhart, J. et al. (2000) Developmental Neurosci. 22: 237
5. Eberhart, J. et al. (2004) J of Neurosci. 24: 1070

View product citations for antibody E254 on CiteAb

CyTOF®

ICC

Certificate of Analysis

Request COA

Prod No.	Description
E254	Anti-Rat EphA5 (Clone 86731)
E260	Anti-Rat EphA5
E289	Anti-Rat EphA5 – Biotin

Formats Available

Prod No.	Description
E254	Anti-Rat EphA5 (Clone 86731)

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Anti-Rat EphA5 (Clone 86731)