Anti-alpha 1 Adrenergic Receptor antibody (ab3462)

Overview

  • Product nameAnti-alpha 1 Adrenergic Receptor antibody
    See all alpha 1 Adrenergic Receptor primary antibodies
  • Description
    Rabbit polyclonal to alpha 1 Adrenergic Receptor
  • SpecificityBy Western blot, this antibody detects a 60 kDa protein representing the A1AR from mouse kidney membrane preparations, mouse liver, HeLa cell lysates, and zebra fish samples. Immunofluorescence staining of A1AR in mouse kidney distal tubule cells results in staining of the plasma membrane.
  • Tested applicationsSuitable for: WB, ELISA, ICC, ICC/IF, IP, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cat, Human, Drosophila melanogaster, Zebrafish
    Predicted to work with: Rabbit, Guinea pig, Hamster, Cow, Dog, Pig, Japanese ricefish
  • Immunogen

    Synthetic peptide corresponding to Human alpha 1 Adrenergic Receptor aa 339-349 (intracellular). Immunizing peptide corresponds to residues in the 3rd intracellular loop of human A1AR. Thiis sequence is 100% conserved in all A1AR subtypes examined.
    Sequence:

    KFSREKKAAKT


    (Peptide available as ab41797)

  • Positive control
    • mouse kidney membrane preparations, mouse liver lysate, HeLa cell lysate

Properties

Applications

Our Abpromise guarantee covers the use of ab3462 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/400. Detects a band of approximately 60 kDa.Can be blocked with alpha 1 Adrenergic Receptor peptide (ab41797).
ELISA Use at an assay dependent concentration.
ICC 1/10 - 1/1000.
ICC/IF 1/1000.
IP Use at an assay dependent concentration.

use 5 ug/mg lysate

IHC-P 1/50 - 1/200.

Target

Anti-alpha 1 Adrenergic Receptor antibody images

  • Immunocytochemistry/Immunofluorescence analysis of alpha 1 Adrenergic Receptor (green) showing staining in the cytoplasm of HepG2 cells (right) compared to a negative control (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were incubated with ab3462 in 3% BSA-PBS at a dilution of 1:100 overnight at 4 ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

  • Immunocytochemistry/Immunofluorescence analysis of alpha 1 Adrenergic Receptor (green) showing staining in the cytoplasm of NIH-3T3 cells (right) compared to a negative control (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were incubated with ab3462 in 3% BSA-PBS at a dilution of 1:100 overnight at 4 ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

  • Immunocytochemistry/Immunofluorescence analysis of alpha 1 Adrenergic Receptor (green) showing staining in the cytoplasm of PC-3 cells (right) compared to a negative control (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were incubated with ab3462 in 3% BSA-PBS at a dilution of 1:100 overnight at 4 ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

  • ab3462 labelling alpha 1 Adrenergic Receptor in the cytoplasm and membrane of Mouse brain tissue (right) compared with a negative control (left). To expose target proteins, antigen retrieval method was performed using 10mM sodium citrate (pH 6.0) microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature. Tissue sections were incubated in the primary antibody (1:100 in 3% BSA-PBS) overnight at 4°C. A HRP-conjugated anti-rabbit was used as the secondary antibody, followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

  • ab3462 labelling alpha 1 Adrenergic Receptor in the cytoplasm and membrane of Human prostate tissue (right) compared with a negative control (left). To expose target proteins, antigen retrieval method was performed using 10mM sodium citrate (pH 6.0) microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature. Tissue sections were incubated in the primary antibody (1:100 in 3% BSA-PBS) overnight at 4°C. A HRP-conjugated anti-rabbit was used as the secondary antibody, followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

  • Immunolocalization of alpha 1 Adrenergic Receptor in mouse distal convoluted tubule using ab3462.

References for Anti-alpha 1 Adrenergic Receptor antibody (ab3462)

This product has been referenced in:
  • Sloniecka M  et al. Expression Profiles of Neuropeptides, Neurotransmitters, and Their Receptors in Human Keratocytes In Vitro and In Situ. PLoS One 10:e0134157 (2015). IHC-Fr, ICC/IF ; Human . Read more (PubMed: 26214847) »
  • Ives SJ  et al. Heat and a1-adrenergic responsiveness in human skeletal muscle feed arteries: the role of nitric oxide. J Appl Physiol 113:1690-8 (2012). WB ; Human . Read more (PubMed: 23042905) »

See all 4 Publications for this product

Product Wall

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample Mouse Tissue sections (heart)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate Buffer
Permeabilization Yes - 0.2% Triton X-100
Specification heart
Blocking step Serum as blocking agent for 10 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Fixative Paraformaldehyde
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Verified customer

Submitted Nov 01 2016

Application Western blot
Sample Mouse Tissue lysate - other (cellular fractionation of hindpaw skin protein)
Gel Running Conditions Reduced Denaturing
Loading amount 50 µg
Specification cellular fractionation of hindpaw skin protein
Blocking step LiCOR blocking buffer as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 50% · Temperature: 24°C
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Submitted Sep 30 2015

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Loading amount 60 µg
Gel Running Conditions Reduced Denaturing (8)
Sample Human Cell lysate - whole cell (MCF7)
Specification MCF7
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
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Submitted Sep 09 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunoprecipitation
Immuno-precipitation step Staph. aureus
Sample Human Cell lysate - whole cell (MCF7)
Specification MCF7
Total protein in input 100 µg
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Submitted Sep 06 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunoprecipitation
Immuno-precipitation step Staph. aureus
Sample Human Cell lysate - whole cell (HELA cells)
Specification HELA cells
Total protein in input 50 µg
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Mr. Bamidipati Naga Krishna Srinivas

Verified customer

Submitted Jun 27 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Loading amount 100 µg
Gel Running Conditions Reduced Denaturing (8)
Sample Human Cell lysate - whole cell (HELA cells)
Specification HELA cells
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
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Submitted Jun 27 2013

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this h...

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Thank you for contacting Abcam.

For the antibody ab3462, I was able to locate WB conditions that were used for testing. Please see below.

The samples tested were HeLa cell lysate. rat liver, mouse liver, drosophila and zebra fish. ...

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I am sorry that you have experienced this error! I would be happy to issue a free of charge replacement of the vial of ab3462 you received that did not contain the full volume. This is a mistake on our part, and I certainly want to ensure that you rece...

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Thank you for your inquiry.

I do not suggest to dilute the antibody in the same buffer it is stored in.
The buffer to create the working solutions should be according to the application the antibody is used.

I cannot recommen...

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