重组Anti-Nogo A + Nogo D抗体[EPR26286-15] - BSA and Azide free (ab318265)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR26286-15] to Nogo A + Nogo D - BSA and Azide free
- Suitable for: ICC/IF, IHC-P, IHC-Fr, WB
- Reacts with: Mouse, Rat
Related conjugates and formulations
概述
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产品名称
Anti-Nogo A + Nogo D抗体[EPR26286-15] - BSA and Azide free
参阅全部 Nogo A+Nogo D 一抗 -
描述
兔单克隆抗体[EPR26286-15] to Nogo A + Nogo D - BSA and Azide free -
宿主
Rabbit -
特异性
Very weak non-specific staining is detected in mouse spleen and kidney tissue, ab315792 is recommended as alternative for mouse IHC.
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经测试应用
适用于: ICC/IF, IHC-P, IHC-Fr, WBmore details -
种属反应性
与反应: Mouse, Rat
不与反应: Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: Mouse brain, Mouse cerebellum, Mouse testis, Rat brain, Rat testis, Purified 293T cells transfected with a mouse Nogo-A expression vector containing a His-tag and Purified 293T cells transfected with a mouse Nogo-D expression vector containing a His-tag lysates. IHC-P: Mouse cerebrum, Mouse spinal cord, Rat cerebrum and Rat spinal cord tissues. IHC-Fr: Rat cerebrum and Mouse cerebrum fresh frozen tissues. ICC/IF: mouse primary neural/glia cell and rat primary neural/glia cells
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常规说明
ab318265 is the carrier-free version of ab318264
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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 production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C. -
存储溶液
pH: 7.2
Constituent: 100% PBS -
无载体
是 -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR26286-15 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab318265于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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ICC/IF |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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IHC-Fr |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration.
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说明 |
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ICC/IF
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
IHC-Fr
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. |
靶标
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细胞定位
Nogo A: Endoplasmic reticulum membrane. Anchored to the membrane of the endoplasmic reticulum through 2 putative transmembrane domains. Nogo D: Endoplasmic reticulum membrane. Anchored to the membrane of the endoplasmic reticulum through 2 putative transmembrane domains. Co-localizes with TMEM33 at the ER sheets. -
数据库链接
- Entrez Gene: 68585 Mouse
- Entrez Gene: 83765 Rat
- SwissProt: Q99P72 Mouse
- SwissProt: Q9JK11 Rat
- Unigene: 192580 Mouse
- Unigene: 440639 Mouse
- Unigene: 445271 Mouse
- Unigene: 472973 Mouse
see all
图片
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All lanes : Anti-Nogo A + Nogo D antibody [EPR26286-15] (ab318264) at 1/1000 dilution
Lane 1 : Mouse brain tissue lysate
Lane 2 : Mouse cerebellum tissue lysate
Lane 3 : Mouse liver tissue lysate
Lane 4 : Mouse testis tissue lysate
Lane 5 : Mouse spleen tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Observed band size: 190 kDa why is the actual band size different from the predicted?This data was developed using ab318264, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Negative control: liver, spleen (PMID: 11978832).
In lane 4-5 lysates were freshly made and used for Western blotting immediately to minimize protein degradation.
The identity of the lower MW band at approximately 80 kDa (in lane 1) is unknown.
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.
Exposure time: Lane 1-3: 48 seconds, Lane 4-5: 180 seconds
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All lanes : Anti-Nogo A + Nogo D antibody [EPR26286-15] (ab318264) at 1/1000 dilution
Lane 1 : Rat brain tissue lysate
Lane 2 : Rat liver tissue lysate
Lane 3 : Rat testis tissue lysate
Lane 4 : Rat spleen tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Observed band size: 190 kDa why is the actual band size different from the predicted?
Exposure time: 180 secondsThis data was developed using ab318264, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Negative control: liver, spleen (PMID: 11978832).
Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.
The identity of the bands between 80 kDa and 120 kDa are unknown.
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.
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All lanes : Anti-Nogo A + Nogo D antibody [EPR26286-15] (ab318264) at 1/1000 dilution
Lane 1 : Purified 293T cells transfected with a mouse Nogo-A expression vector containing a His-tag, whole cell lysate
Lane 2 : Purified 293T cells transfected with a mouse Nogo-B2 expression vector containing a His-tag, whole cell lysate
Lane 3 : Purified 293T cells transfected with a mouse Nogo-C expression vector containing a His-tag, whole cell lysate
Lane 4 : Purified 293T cells transfected with a mouse Nogo-D expression vector containing a His-tag, whole cell lysate
Lane 5 : Purified 293T cells transfected with a mouse Nogo-B1 expression vector containing a His-tag, whole cell lysate
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution
Observed band size: 190 kDa why is the actual band size different from the predicted?
Exposure time: 180 secondsThis data was developed using ab318264, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
This antibody cross-react with mouse Nogo D, does not cross-react with mouse Nogo-B2, Nogo-C and Nogo-B1.
In Western blot, Anti-6X His tag antibody [EPR20547]-CHIP Grade (ab213204) staining at 1/5000 dilution.
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This data was developed using ab318264, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling Nogo A + Nogo D with ab318264 at 1/5000 (0.099 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond Polymer Refine Detection).
Positive staining in mouse cerebrum.
The section was incubated with ab318264 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND ® RX instrumentCounterstained 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins
-
This data was developed using ab318264, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Mouse spinal cord tissue labeling Nogo A + Nogo D with ab318264 at 1/5000 (0.099 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond Polymer Refine Detection).
Positive staining in mouse spinal cord.
The section was incubated with ab318264 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND ® RX instrumentCounterstained 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins
-
This data was developed using ab318264, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling Nogo A + Nogo D with ab318264 at 1/5000 (0.099 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond Polymer Refine Detection).
Positive staining in rat cerebrum.
The section was incubated with ab318264 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND ® RX instrumentCounterstained 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins
-
This data was developed using ab318264, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Rat spinal cord tissue labeling Nogo A + Nogo D with ab318264 at 1/5000 (0.099 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond Polymer Refine Detection).
Positive staining in rat spinal cord.
The section was incubated with ab318264 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND ® RX instrumentCounterstained 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins
-
This data was developed using ab318264, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Nogo A + Nogo D with ab318264 at 1/5000 (0.099 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond Polymer Refine Detection).
Negative control: No staining in mouse liver.
The section was incubated with ab318264 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND ® RX instrumentCounterstained 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins
-
This data was developed using ab318264, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling Nogo A + Nogo D with ab318264 at 1/5000 (0.099 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond Polymer Refine Detection).
Negative control: No staining in rat liver.
The section was incubated with ab318264 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND ® RX instrumentCounterstained 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins
-
This data was developed using ab318264, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat cerebrum (fresh frozen) tissue labeling Nogo A + Nogo D with ab318264 at 1/500 (0.992 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor ® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).
Confocal image showing positive staining on rat cerebrum. The nuclear counterstain was DAPI (Blue). The section was incubated with ab318264 for 60 mins at room temperature. The section was then mounted using Fluoromount ®.The immunostaining was performed on a Leica Biosystems BOND ® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Secondary antibody control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor ® 488) preadsorbedat 1/1000 2 ug/mL dilution.
-
This data was developed using ab318264, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat liver (fresh frozen) tissue labeling Nogo A + Nogo D with ab318264 at 1/500 (0.992 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor ® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).
Negative control: confocal image showing no staining on rat liver. The nuclear counterstain was DAPI (Blue). The section was incubated with ab318264 for 60 mins at room temperature. The section was then mounted using Fluoromount ®.The immunostaining was performed on a Leica Biosystems BOND ® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Secondary antibody control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor ® 488) preadsorbedat 1/1000 2 ug/mL dilution.
-
This data was developed using ab318264, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse cerebrum (fresh frozen) tissue labeling Nogo A + Nogo D with ab318264 at 1/500 (0.992 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor ® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).
Confocal image showing positive staining on mouse cerebrum. The nuclear counterstain was DAPI (Blue). The section was incubated with ab318264 for 60 mins at room temperature. The section was then mounted using Fluoromount ®.The immunostaining was performed on a Leica Biosystems BOND ® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Secondary antibody control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor ® 488) preadsorbedat 1/1000 2 ug/mL dilution.
-
This data was developed using ab318264, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse liver (fresh frozen) tissue labeling Nogo A + Nogo D with ab318264 at 1/500 (0.992 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor ® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).
Negative control: confocal image showing no staining on mouse liver. The nuclear counterstain was DAPI (Blue). The section was incubated with ab318264 for 60 mins at room temperature. The section was then mounted using Fluoromount ®.The immunostaining was performed on a Leica Biosystems BOND ® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Secondary antibody control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor ® 488) preadsorbedat 1/1000 2 ug/mL dilution.
-
This data was developed using ab318264, the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neural/glia cell cells labelling Nogo A + Nogo D with ab318264 at 1/50 (9.92 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 mouse primary neural/glia cell (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection.
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 4ug/ml dilution (Magenta). 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.
-
This data was developed using ab318264, the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse splenocytes cells labelling Nogo A + Nogo D with ab318264 at 1/50 (9.92 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).
Negative control: Confocal image showing negative staining in mouse splenocytes (PMID: 11978832) (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
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 2.5ug/ml dilution (Magenta). 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.
-
This data was developed using ab318264, the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized rat primary neural/glia cell cells labelling Nogo A + Nogo D with ab318264 at 1/50 (9.92 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 rat primary neural/glia cell (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection.
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 4ug/ml dilution (Magenta). 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.
-
This data was developed using ab318264, the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized rat splenocytes cells labelling Nogo A + Nogo D with ab318264 at 1/50 (9.92 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).
Negative control: Confocal image showing negative staining in rat splenocytes (PMID: 11978832) (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
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 2.5ug/ml dilution (Magenta). 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.
实验方案
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
数据表及文件
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Datasheet download
Certificate of Compliance
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