Anti-Dopamine D1a Receptor (mouse monoclonal)

Species Reactivity	Key Applications	Host	Format	Antibody Type
M, R, Sh	IHC, WB	M	Purified	Monoclonal Antibody

Species Reactivity

Key Applications

Host

Format

Antibody Type

M, R, Sh

IHC, WB

Purified

Monoclonal Antibody

Description
Catalogue Number	MAB5290
Brand Family	Chemicon®
Trade Name	Chemicon
Description	Anti-Dopamine D1A Receptor Antibody, clone SG2-D1a

Description

Catalogue Number

MAB5290

Brand Family

Chemicon®

Trade Name

Chemicon

Description

Anti-Dopamine D1A Receptor Antibody, clone SG2-D1a

Product Information
Format	Purified
Presentation	Purified immunoglobulin. Liquid in 0.02 M Phosphate buffer, 0.25 M NaCl, pH 7.6 with 0.1% sodium azide.

Product Information

Format

Purified

Presentation

Purified immunoglobulin. Liquid in 0.02 M Phosphate buffer, 0.25 M NaCl, pH 7.6 with 0.1% sodium azide.

Applications
Application	Anti-Dopamine D1A Receptor Antibody, clone SG2-D1a detects level of Dopamine D1A Receptor & has been published & validated for use in IH & WB.
Key Applications	Immunohistochemistry Western Blotting
Application Notes	Western blot: 1:250-1:500 on unboiled membrane fractions from rat caudate tissue. Reactivity is also seen in perfrontal and temporal cortex. An identified band between 45-66kDa is observed (Luedtke, 1999). Immunohistochemistry: 4% PFA fixed, vibratome sections, blocked with 3% NGS, 1% BSA with 0.2% triton X-100. Primary antibodies diluted onlyin PBS with excess blocking reagents. Optimal working dilutions must be determined by the end user. SUGGESTED WESTERN BLOT PROTOCOL Solutions 10X Transfer buffer 1 liter Tris 115.3 gm Glycine 24.2 gm [SDS 4 gm (may be omitted)] For a 1X working solution: 1 liter mix 700 mL water 200 mL MeOH 100 mL 10X TransferTBST stock 1 liter 1M Tris pH 7.5 10 mL 5 M NaC 30 mL 10% Tween-20 5 mL 1. Run SDS-PAGE gel as desired. 2. Soak PVDF membrane (ImmobilonP from Millipore) in 100% methanol for 1-2 minutes to wet the membrane. 3. Soak membrane in transfer buffer (see below) until ready to set up blot transfer (>15 min). 4. Assemble gel-transfer sandwich with the blot toward the positive electrode. 5. Transfer protein from gel to membrane at 100 volts for 1-2 hours for small molecular weight proteins or 65 mA for 16 hours (overnight) for complete transfer of higher molecular weight proteins. 6. Stain the transferred bands with Chemicon BLOT-FastStain™ (Catalog Number 2076). 7. Destain with deionized water. 8. Block with 5% non-fat milk (Marvel or Carnation) in TBS or PBS, overnight at 4°C. The non-fat milk should be dissolved freshly, centrifuged 10,000 rpm for 10 min, and filtered through glass filter (Gelman Acrodisc). 9. Incubation with first antibody overnight with gentle agitation at 4°C. Optimal working dilutions and incubation time will need to be determined by the end user. 10. Wash at least 3 x 5 min. with TBST. From this stage, azide should be omitted. 11. Incubation with the secondary antibody (HRP-conjugated goat anti-mouse antibody, for example Chemicon Catalog Number AP124P, diluted appropriately) 1 h at room temperature. 12. Wash at least 3 x 5 min. with TBST. 13. Perform ECL with commercial kit (femtoLUCENT, Chemicon Catalog Number 2078).

Applications

Application

Anti-Dopamine D1A Receptor Antibody, clone SG2-D1a detects level of Dopamine D1A Receptor & has been published & validated for use in IH & WB.

Key Applications

Immunohistochemistry
Western Blotting

Application Notes

Western blot: 1:250-1:500 on unboiled membrane fractions from rat caudate tissue. Reactivity is also seen in perfrontal and temporal cortex. An identified band between 45-66kDa is observed (Luedtke, 1999).

Immunohistochemistry: 4% PFA fixed, vibratome sections, blocked with 3% NGS, 1% BSA with 0.2% triton X-100. Primary antibodies diluted onlyin PBS with excess blocking reagents.

Optimal working dilutions must be determined by the end user.

SUGGESTED WESTERN BLOT PROTOCOL

Solutions

10X Transfer buffer 1 liter

Tris 115.3 gm

Glycine 24.2 gm

[SDS 4 gm (may be omitted)]

For a 1X working solution: 1 liter

mix 700 mL water

200 mL MeOH

100 mL 10X TransferTBST

stock 1 liter

1M Tris pH 7.5 10 mL

5 M NaC 30 mL

10% Tween-20 5 mL

1. Run SDS-PAGE gel as desired.

2. Soak PVDF membrane (ImmobilonP from Millipore) in 100% methanol for 1-2 minutes to wet the membrane.

3. Soak membrane in transfer buffer (see below) until ready to set up blot transfer (>15 min).

4. Assemble gel-transfer sandwich with the blot toward the positive electrode.

5. Transfer protein from gel to membrane at 100 volts for 1-2 hours for small molecular weight proteins or 65 mA for 16 hours (overnight) for complete transfer of higher molecular weight proteins.

6. Stain the transferred bands with Chemicon BLOT-FastStain™ (Catalog Number 2076).

7. Destain with deionized water.

8. Block with 5% non-fat milk (Marvel or Carnation) in TBS or PBS, overnight at 4°C. The non-fat milk should be dissolved freshly, centrifuged 10,000 rpm for 10 min, and filtered through glass filter (Gelman Acrodisc).

9. Incubation with first antibody overnight with gentle agitation at 4°C.

Optimal working dilutions and incubation time will need to be determined by the end user.

10. Wash at least 3 x 5 min. with TBST. From this stage, azide should be omitted.

11. Incubation with the secondary antibody (HRP-conjugated goat anti-mouse antibody, for example Chemicon Catalog Number AP124P, diluted appropriately) 1 h at room temperature.

12. Wash at least 3 x 5 min. with TBST.

13. Perform ECL with commercial kit (femtoLUCENT, Chemicon Catalog Number 2078).

Biological Information
Immunogen	Recombinant rat D1a Receptor, carboxy terminus.
Clone	SG2-D1a
Concentration	Please refer to the Certificate of Analysis for the lot-specific concentration.
Host	Mouse
Specificity	Dopamine Receptor D1A, no cross reactivity to D1B (D5) receptor. Immunoreactivity was found to be specific for mouse and rat D1A receptor. Human, pig, dog, rabbit, sheep and bovine samples were non-reactive by western analysis.
Isotype	IgG2b
Species Reactivity	Mouse Rat Sheep
Antibody Type	Monoclonal Antibody
Entrez Gene Number	NM_000794.3
Entrez Gene Summary	This gene encodes the D1 subtype of the dopamine receptor. The D1 subtype is the most abundant dopamine receptor in the central nervous system. This G-protein coupled receptor stimulates adenylyl cyclase and activates cyclic AMP-dependent protein kinases. D1 receptors regulate neuronal growth and development, mediate some behavioral responses, and modulate dopamine receptor D2-mediated events. Alternate transcription initiation sites result in two transcript variants of this gene.
Gene Symbol	DRD1 DRD1A DADR D
Non-Reactive Species	Bovine Canine Human Pig Rabbit
UniProt Number	P21728
UniProt Summary	FUNCTION: SwissProt: P21728 # This is one of the five types (D1 to D5) of receptors for dopamine. The activity of this receptor is mediated by G proteins which activate adenylyl cyclase. SIZE: 446 amino acids; 49293 Da SUBUNIT: Interacts with DNAJC14 via its C-terminus (By similarity). Interacts with DRD1IP. SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein (By similarity). Endoplasmic reticulum membrane; Multi-pass membrane protein (By similarity). Note=Transport from the endoplasmic reticulum to the cell surface is regulated by interaction with DNAJC14 (By similarity). TISSUE SPECIFICITY: Detected in caudate, nucleus accumbens and in the olfactory tubercle. SIMILARITY: SwissProt: P21728 ## Belongs to the G-protein coupled receptor 1 family.

Biological Information

Immunogen

Recombinant rat D1a Receptor, carboxy terminus.

Clone

SG2-D1a

Concentration

Please refer to the Certificate of Analysis for the lot-specific concentration.

Host

Mouse

Specificity

Dopamine Receptor D1A, no cross reactivity to D1B (D5) receptor. Immunoreactivity was found to be specific for mouse and rat D1A receptor. Human, pig, dog, rabbit, sheep and bovine samples were non-reactive by western analysis.

Isotype

IgG2b

Species Reactivity

Mouse
Rat
Sheep

Antibody Type

Monoclonal Antibody

Entrez Gene Number

NM_000794.3

Entrez Gene Summary

This gene encodes the D1 subtype of the dopamine receptor. The D1 subtype is the most abundant dopamine receptor in the central nervous system. This G-protein coupled receptor stimulates adenylyl cyclase and activates cyclic AMP-dependent protein kinases. D1 receptors regulate neuronal growth and development, mediate some behavioral responses, and modulate dopamine receptor D2-mediated events. Alternate transcription initiation sites result in two transcript variants of this gene.

Gene Symbol

DRD1
DRD1A
DADR
D

Non-Reactive Species

Bovine
Canine
Human
Pig
Rabbit

UniProt Number

P21728

UniProt Summary

FUNCTION: SwissProt: P21728 # This is one of the five types (D1 to D5) of receptors for dopamine. The activity of this receptor is mediated by G proteins which activate adenylyl cyclase.
SIZE: 446 amino acids; 49293 Da
SUBUNIT: Interacts with DNAJC14 via its C-terminus (By similarity). Interacts with DRD1IP.
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein (By similarity). Endoplasmic reticulum membrane; Multi-pass membrane protein (By similarity). Note=Transport from the endoplasmic reticulum to the cell surface is regulated by interaction with DNAJC14 (By similarity).
TISSUE SPECIFICITY: Detected in caudate, nucleus accumbens and in the olfactory tubercle.
SIMILARITY: SwissProt: P21728 ## Belongs to the G-protein coupled receptor 1 family.

Product Usage Statements
Usage Statement	Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Product Usage Statements

Usage Statement

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Storage and Shipping Information
Storage Conditions	Maintain at 2-8°C in undiluted aliquots for up to 6 months after date of receipt.

Storage and Shipping Information

Storage Conditions

Maintain at 2-8°C in undiluted aliquots for up to 6 months after date of receipt.

Packaging Information
Material Size	100 µg

Packaging Information

Material Size

100 µg

实验耗材

分子生物学试剂

蛋白与免疫学

生化试剂

细胞及检测试剂

色谱及层析

仪器设备

实验服务

办公用品

Anti-Dopamine D1a Receptor (mouse monoclonal)

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