SHANK1+SHANK3+SHANK2重组抗体[EPR26549-87]_SHANK1+SHANK3+SHANK2抗体(ab317608)

产品名称

Anti-SHANK1 + SHANK2 + SHANK3抗体[EPR26549-87]
描述

兔单克隆抗体[EPR26549-87] to SHANK1 + SHANK3 + SHANK2
宿主

Rabbit
特异性

Unsuitable for rat IHC-Fr and human IHC-P.
经测试应用

适用于: IHC-P, IHC-Fr, WB, Flow Cyt (Intra)more details
不适用于: ICC/IF or IP
种属反应性

与反应: Mouse, Rat, Human
免疫原

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
阳性对照
- WB: Human hippocampus,T-47D, Mouse cerebellum, Mouse hippocampus, Rat cerebellum, Rat hippocampus, His-tagged human SHANK2 recombinant fragment, His-tagged human SHANK1 recombinant fragment and His-tagged human SHANK3 recombinant fragment lysates. IHC-P: Mouse liver, Mouse hippocampus, Rat liver, Rat cerebellum tissues. IHC-Fr: Mouse hippocampus (fresh frozen) tissues. Flow Cyt (Intra): T-47D, 293T, Mouse primary neuron cells.
常规说明
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

形式

Liquid
存放说明

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
存储溶液

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 59% PBS, 0.05% BSA
Concentration information loading...
纯度

Protein A purified
克隆

单克隆
克隆编号

EPR26549-87
同种型

IgG
研究领域
- Neuroscience
- Processes

Alternative Versions
- Anti-SHANK1 + SHANK2 + SHANK3 antibody [EPR26549-87] - BSA and Azide free (ab317609)
Compatible Secondaries
- Rabbit specific IHC polymer detection kit HRP/DAB (ab209101)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Related Products

The Abpromise guarantee

Abpromise™承诺保证使用ab317608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
IHC-P		1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IHC-Fr		1/100.
WB		1/1000.
Flow Cyt (Intra)		1/500.

说明
IHC-P 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IHC-Fr 1/100.
WB 1/1000.
Flow Cyt (Intra) 1/500.

应用说明

Is unsuitable for ICC/IF or IP.

细胞定位

SHANK1: Cytoplasm. Cell junction > synapse > postsynaptic cell membrane > postsynaptic density. Cell junction > synapse. Postsynaptic density of neuronal cells. SHANK3: Cytoplasm. Cell junction > synapse. Cell junction > synapse > postsynaptic cell membrane > postsynaptic density. Postsynaptic density of neuronal cells. SHANK2: Apical cell membrane. Cytoplasm. Cell junction, synapse. Cell junction, synapse, postsynaptic cell membrane, postsynaptic density. Cell projection, growth cone. Cell projection, dendritic spine. Colocalizes with cortactin in growth cones in differentiating hippocampal neurons. Present in the dendritic spines of cerebellar Purkinje cells (By similarity). Colocalizes with cortactin in growth cones in differentiating hippocampal neurons. Colocalized with PDE4D to the apical membrane of colonic crypt cells.
数据库链接
- Entrez Gene: 22941 Human
- Entrez Gene: 50944 Human
- Entrez Gene: 85358 Human
- Entrez Gene: 210274 Mouse
- Entrez Gene: 243961 Mouse
- Entrez Gene: 58234 Mouse
- Entrez Gene: 171093 Rat
- Entrez Gene: 59312 Rat
- Entrez Gene: 78957 Rat
- Omim: 603290 Human
- Omim: 604999 Human
- Omim: 606230 Human
- SwissProt: Q9BYB0 Human
- SwissProt: Q9UPX8 Human
- SwissProt: Q9Y566 Human
- SwissProt: D3YZU1 Mouse
- SwissProt: Q4ACU6 Mouse
- SwissProt: Q80Z38 Mouse
- SwissProt: Q9JLU4 Rat
- SwissProt: Q9QX74 Rat
- SwissProt: Q9WV48 Rat
- Unigene: 149035 Human
- Unigene: 268726 Human
- Unigene: 274255 Human
- Unigene: 146855 Mouse
- Unigene: 360368 Mouse
- Unigene: 483624 Mouse
- Unigene: 14545 Rat
- Unigene: 225968 Rat
- Unigene: 42876 Rat
see all

Western blot - Anti-SHANK1 + SHANK2 + SHANK3 antibody [EPR26549-87] (ab317608)

All lanes : Anti-SHANK1 + SHANK2 + SHANK3 antibody [EPR26549-87] (ab317608) at 1/1000 dilution

Lane 1 : Human hippocampus tissue lysate
Lane 2 : Human skeletal muscle tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/200000 dilution

Observed band size: 180-240 kDa why is the actual band size different from the predicted?

Exposure time: 180 seconds

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

Negative control: skeletal muscle (PMID: 10506216; PMID: 22346768).

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID:10506216). The bands beneath the target band (180-240 kDa) are likely to be degraded target fragments.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.
Western blot - Anti-SHANK1 + SHANK2 + SHANK3 antibody [EPR26549-87] (ab317608)

All lanes : Anti-SHANK1 + SHANK2 + SHANK3 antibody [EPR26549-87] (ab317608) at 1/1000 dilution

Lane 1 : T-47D (human ductal breast epithelial tumor epithelial cell) whole cell lysate
Lane 2 : 293T (human embryonic kidney epithelial cell) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Observed band size: 180-270 kDa why is the actual band size different from the predicted?

Exposure time: 15 seconds

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

Low expression: 293T (PMID:32661924).

The molecular weight observed is consistent with what has been described in the literature (PMID:10506216). The bands beneath the target band (180-270 kDa) are likely to be degraded target fragments.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.
Western blot - Anti-SHANK1 + SHANK2 + SHANK3 antibody [EPR26549-87] (ab317608)

All lanes : Anti-SHANK1 + SHANK2 + SHANK3 antibody [EPR26549-87] (ab317608) at 1/1000 dilution

Lane 1 : Mouse cerebellum tissue lysate
Lane 2 : Mouse hippocampus tissue lysate
Lane 3 : Mouse skeletal muscle tissue lysate
Lane 4 : Rat cerebellum tissue lysate
Lane 5 : Rat hippocampus tissue lysate
Lane 6 : Rat skeletal muscle tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Observed band size: 180-270 kDa why is the actual band size different from the predicted?

Exposure time: 59 seconds

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

Negative control: skeletal muscle (PMID: 10506216; PMID: 22346768).

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID:10506216). The bands beneath the target band (180-270 kDa) are likely to be degraded target fragments.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.
Western blot - Anti-SHANK1 + SHANK2 + SHANK3 antibody [EPR26549-87] (ab317608)

Lanes 1-2 : Anti-SHANK1 + SHANK2 + SHANK3 antibody [EPR26549-87] (ab317608) at 1/1000 dilution

Lane 1 : His-tagged human SHANK2 recombinant fragment
Lane 2 : His-tagged human SHANK1 recombinant fragment
Lane 3 : His-tagged human SHANK3 recombinant fragment

Lysates/proteins at 0.01 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Observed band size: 26 kDa why is the actual band size different from the predicted?

Exposure time: 125 seconds

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

This antibody can recognize human SHANK1, SHANK2 and SHANK3. .

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHANK1 + SHANK2 + SHANK3 antibody [EPR26549-87] (ab317608)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling SHANK1 + SHANK2 + SHANK3 with ab317608 at 1/100 (4.75 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on cholangiocytes in mouse liver (PMID: 14977424). The section was incubated with ab317608 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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-SHANK1 + SHANK2 + SHANK3 antibody [EPR26549-87] (ab317608)

Immunohistochemical analysis of paraffin-embedded Mouse hippocampus tissue labeling SHANK1 + SHANK2 + SHANK3 with ab317608 at 1/100 (4.75 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining in mouse hippocampus (PMID: 27581745). The section was incubated with ab317608 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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-SHANK1 + SHANK2 + SHANK3 antibody [EPR26549-87] (ab317608)

Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling SHANK1 + SHANK2 + SHANK3 with ab317608 at 1/100 (4.75 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on cholangiocytes in rat liver (PMID: 14977424). The section was incubated with ab317608 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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-SHANK1 + SHANK2 + SHANK3 antibody [EPR26549-87] (ab317608)

Immunohistochemical analysis of paraffin-embedded Rat cerebellum tissue labeling SHANK1 + SHANK2 + SHANK3 with ab317608 at 1/100 (4.75 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining in rat cerebellum (PMID: 27581745). The section was incubated with ab317608 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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-SHANK1 + SHANK2 + SHANK3 antibody [EPR26549-87] (ab317608)

Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling SHANK1 + SHANK2 + SHANK3 with ab317608 at 1/100 (4.75 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Negative control: No staining on mouse skeletal muscle (PMID: 10506216). The section was incubated with ab317608 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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-SHANK1 + SHANK2 + SHANK3 antibody [EPR26549-87] (ab317608)

Immunohistochemical analysis of paraffin-embedded Rat skeletal muscle tissue labeling SHANK1 + SHANK2 + SHANK3 with ab317608 at 1/100 (4.75 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Negative control: No staining on rat skeletal muscle (PMID: 10506216). The section was incubated with ab317608 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
Immunohistochemistry (Frozen sections) - Anti-SHANK1 + SHANK2 + SHANK3 antibody [EPR26549-87] (ab317608)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse hippocampus (fresh frozen) tissue labeling SHANK1 + SHANK2 + SHANK3 with ab317608 at 1/100 (4.75 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 hippocampus. The nuclear counterstain was DAPI (Blue). The section was incubated with ab317608 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.
Immunohistochemistry (Frozen sections) - Anti-SHANK1 + SHANK2 + SHANK3 antibody [EPR26549-87] (ab317608)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse skeletal muscle (fresh frozen) tissue labeling SHANK1 + SHANK2 + SHANK3 with ab317608 at 1/100 (4.75 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 skeletal muscle. The nuclear counterstain was DAPI (Blue). The section was incubated with ab317608 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.
Flow Cytometry (Intracellular) - Anti-SHANK1 + SHANK2 + SHANK3 antibody [EPR26549-87] (ab317608)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized T-47D (human ductal breast epithelial tumor epithelial cell) (Right) / 293T (human embryonic kidney epithelial cell) (Left) cells labelling SHANK1 + SHANK2 + SHANK3 with ab317608 at 1/500 dilution (0.1ug)/Red (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).
Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.
Low expression: 293T (PMID: 32661924)
Flow Cytometry (Intracellular) - Anti-SHANK1 + SHANK2 + SHANK3 antibody [EPR26549-87] (ab317608)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Mouse primary neuron cells labelling SHANK1 + SHANK2 + SHANK3 with ab317608 at 1/500 dilution (0.1ug)/right (Red) compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).
Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.
Flow Cytometry (Intracellular) - Anti-SHANK1 + SHANK2 + SHANK3 antibody [EPR26549-87] (ab317608)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Mouse spleen cell cells labelling SHANK1 + SHANK2 + SHANK3 with ab317608 at 1/500 dilution (0.1ug)/right (Red) compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).
Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.
Negative control: mouse spleen cell.
Anti-SHANK1 + SHANK2 + SHANK3 antibody [EPR26549-87] (ab317608)

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.

Click here to view the general protocols

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概述

产品名称

描述

宿主

特异性

经测试应用

种属反应性

免疫原

阳性对照

常规说明

性能

形式

存放说明

存储溶液

纯度

克隆

克隆编号

同种型

研究领域

相关产品

Alternative Versions

Compatible Secondaries

Isotype control

Related Products

应用

The Abpromise guarantee

靶标

细胞定位

数据库链接

图片

实验方案

数据表及文件

SDS download

Datasheet download

Certificate of Compliance

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