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Anti-5HT2A Receptor antibody (ab66049)

Submit a review Q&A (2)References (13)
Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-5HT2A Receptor antibody (ab66049)
  • Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-5HT2A Receptor antibody (ab66049)

Key features and details

  • Rabbit polyclonal to 5HT2A Receptor
  • Suitable for: IHC-FoFr
  • Reacts with: Mouse, Rat, Human
  • Isotype: IgG

Overview

  • Product name

    Anti-5HT2A Receptor antibody
    See all 5HT2A Receptor primary antibodies

  • Description

    Rabbit polyclonal to 5HT2A Receptor

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-FoFrmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide corresponding to Rat 5HT2A Receptor aa 1-100 (N terminal).

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

  • Positive control

    • Rat cortex, amygdala and hippocampus tissue; mouse frontal cortical neuron cultures.

  • General notes

    Upon delivery dilute with phosphate buffer or Tris buffer at dilutions no higher than 1/10, aliquot and freeze at -15° C or lower. Antibody can be stored for up to six months if handled as described above.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab66049 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
IHC-FoFr
1/300 - 1/500. Biotin/Streptavidin-HRP Technique.
Notes
IHC-FoFr
1/300 - 1/500. Biotin/Streptavidin-HRP Technique.

Target

  • Function

    This is one of the several different receptors for 5-hydroxytryptamine (serotonin), a biogenic hormone that functions as a neurotransmitter, a hormone, and a mitogen. This receptor mediates its action by association with G proteins that activate a phosphatidylinositol-calcium second messenger system. This receptor is involved in tracheal smooth muscle contraction, bronchoconstriction, and control of aldosterone production.

  • Sequence similarities

    Belongs to the G-protein coupled receptor 1 family.

  • Domain

    The PDZ domain-binding motif is involved in the interaction with INADL, CASK, APBA1, DLG1 and DLG4.

  • Cellular localization

    Cell membrane. Localizes to the post-synaptic thickening of axo-dendritic synapses.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • 5 HT 2 antibody
    • 5 HT 2A antibody
    • 5 HT2 receptor antibody
    • 5 HT2A antibody
    • 5 hydroxytryptamine receptor 2A antibody
    • 5-HT-2 antibody
    • 5-HT-2A antibody
    • 5-hydroxytryptamine (serotonin) receptor 2A, G protein-coupled antibody
    • 5-hydroxytryptamine 2A receptor antibody
    • 5-hydroxytryptamine receptor 2A antibody
    • 5HT2A_HUMAN antibody
    • HTR 2 antibody
    • HTR 2A antibody
    • HTR2 antibody
    • HTR2, formerly antibody
    • HTR2A antibody
    • serotonin 5-HT-2 receptor, formerly antibody
    • serotonin 5-HT-2A receptor antibody
    • Serotonin receptor 2A antibody
    see all

Images

  • Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-5HT2A Receptor antibody (ab66049)
    Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-5HT2A Receptor antibody (ab66049)
    ab66049 at 1/100 dilution staining 5HT2A Receptor in Mouse frontal cortical neuron cultures (p1 pups, day 4 culture, for 48hrs) by Immunohistochemistry, Paraformaldehyde-fixed tissue.
  • Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-5HT2A Receptor antibody (ab66049)
    Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-5HT2A Receptor antibody (ab66049)
    ab66049, at 1/300 dilution staining 5HT2A Receptor in Rat cerebral cortex by Immunohistochemistry, Paraformaldehyde-fixed tissue.

References (13)

Publishing research using ab66049? Please let us know so that we can cite the reference in this datasheet.

ab66049 has been referenced in 13 publications.

  • Ju A  et al. Expression of serotonin 1A and 2A receptors in molecular- and projection-defined neurons of the mouse insular cortex. Mol Brain 13:99 (2020). PubMed: 32594910
  • Gao W  et al. HTR2A promotes the development of cardiac hypertrophy by activating PI3K-PDK1-AKT-mTOR signaling. Cell Stress Chaperones 25:899-908 (2020). PubMed: 32519137
  • Eickelbeck D  et al. CaMello-XR enables visualization and optogenetic control of Gq/11 signals and receptor trafficking in GPCR-specific domains. Commun Biol 2:60 (2019). PubMed: 30793039
  • Delicata F  et al. Preferential modulation of the lateral habenula activity by serotonin-2A rather than -2C receptors: Electrophysiological and neuroanatomical evidence. CNS Neurosci Ther 24:721-733 (2018). PubMed: 29479825
  • Szlachta M  et al. Repeated Clozapine Increases the Level of Serotonin 5-HT1AR Heterodimerization with 5-HT2Aor Dopamine D2Receptors in the Mouse Cortex. Front Mol Neurosci 11:40 (2018). PubMed: 29497362
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-2 of 2 Abreviews or Q&A

Question

This customer has purchased ab66049 (Anti-5HT2A Receptor antibody) and has conducted the wb several times with rat sample. The results show no major band and high background, therefore, he would like to ask for your help to modify his experiment step, could you please offer any suggestion to improve his result?

I attached the image in this letter and her experiment step as follow:



1. Order details:


Batch number: gr55638-1

Po: 1028918

Abcam product code: ab66049

Antibody storage conditions (temperature/reconstitution etc) aliquot and stored at -20oC




2. Please describe the problem (high background, wrong band size, more bands, no band etc).

No major band and high background

3. On what material are you testing the antibody in WB?

· Species: SD rat, female

· What’s cell line or tissue: brain tissue; hippocampus & prefrontal cortex

· Cell extract or Nuclear extract:

· Purified protein or Recombinant protein: Purified



3. The lysate


How much protein was loaded: 10 microgram

What lysis buffer was used: self-made (sucrose, EDTA, and protease inhibitor cocktail)



What protease inhibitors were used: sigma (SI-P2714-1BTL)

What loading buffer was used: buffer was prepared with the following components of Tris-HCl (pH6.8), SDS, glycerol, beta-mercaptoethanol, EDTA, bromophenol blue

Phosphatase inhibitors

Did you heat the samples: temperature and time: 95oC for 5minutes, followed by 4oC-incubation for 3 mintues




4. Electrophoresis/Gel conditions/ Transfer conditions


Reducing or non reducing gel: reducing

Reducing agent: beta-mercaptoethanol

Gel percentage : 8% running gel

Transfer conditions: (Type of membrane, Protein transfer verified): PVDF, protein marker was completely transferred




5. Blocking conditions


Buffer: TBST

Blocking agent: milk, BSA, serum, what percentage: milk, 5%

Incubation time: 1 hour

Incubation temperature: room temperature




6. Primary Antibody


Species:

Reacts against:


· At what dilution(s) have you tested this antibody:

· What dilution buffer was used:

· Incubation time:

· Incubation temperature:

· What washing steps were done:



7. Secondary Antibody


Species:Rabbit

Reacts against: mouse and rat

At what dilution(s) have you tested this antibody: 1/100 as suggested

Incubation time: over 14 hours at 4oC

Wash steps: membrane soaked in TBST and incubated in gentle shake for 5 minutes at room temperature; 3 times

Fluorochrome or enzyme conjugate: enzyme conjugate; HRP

Do you know whether the problems you are experiencing come from the secondary? Yes, and I see no problem with the secondary since it was applied on other antibody detection on the same membrane, and they all worked out, except anti-5HT 2A




8. Detection method
ECl, ECl+, other detection method: ECL



9. Did you apply positive and negative controls along with the samples? Please specify.



I only got the internal control (beta actin) for quantitative analysis

10. Optimization attempts

· How many times have you tried the Western? 3

· Have you eliminated the possibility that any background bands could be due to the secondary antibody? (Run a “No primary” control): Yes, since there was no any band found

· Do you obtain the same results every time e.g. are background bands always in the same place? The background was clear

· What steps have you altered? Primary antibody dilution



Could you please help this customer to solve the problem?

Thanks for your kindly help

Read More
Answer

Thank you for taking time to complete our questionnaire. I am sorry to hear that this antibody is not providing satisfactory results.

The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality.

Having reviewed this case, I would like to offer some suggestions to help optimize the results from abab66049 Anti-5HT2A Receptor antibody. I would also appreciate if you can confirm some further details:


1. As this is a multipass membrane protein, which tent to aggregate in the loading buffer when boiled at high temperature due to their hydrophobic nature, I would like to recommend lowering the boiling temperature down to 60C and prolonging the boiling time up to 10 min. In addition, use DTT instead of beta-mercaptoethanol as this kind of aggregation is favoured by b- ME.

2. Actin is a reliable control for cytoplasmic proteins, but as you are searching for an membrane protein, I would suggest to use an appropriate membrane control.

3. In order to avoid under-loading, I would like to suggest loading 20 to 30 ug protein per lane.

4. Unfortunately, I could not find any information regarding the primaryantibody concentration and incubation set up. My guess is that this information can be found in the Secondary Antibody section. Could you please check this?

5.Have you stripped the membrane before you applied this antibody? If so, how often?

6. I am a bit confused regarding the secondary antibody: From how I understand it, you are using a rabbit secondary antibody against mouse andrat. Could you please confirm in which species the secondary antibody was produced and which species it detects?


In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund.

I thank you for your cooperation.

Read More

Question

To whom it may concern,
1. Order details:
- Batch number:
- Abcam product code: ab66049
- Antibody storage conditions (temperature/reconstitution etc) aliquot and stored at -20oC
2. Please describe the problem (high background, wrong band size, more bands, no band etc).
No band or smear
3. On what material are you testing the antibody in WB?
- Species: SD rat, female
- What’s cell line or tissue: brain tissue; hippocampus & prefrontal cortex
- Cell extract or Nuclear extract:
- Purified protein or Recombinant protein: Purified
3. The lysate
- How much protein was loaded: 10 microgram
- What lysis buffer was used: self-made (sucrose, EDTA, and protease inhibitor cocktail)
- What protease inhibitors were used: sigma (SI-P2714-1BTL)
- What loading buffer was used: buffer was prepared with the following components of Tris-HCl (pH6.8), SDS, glycerol, beta-mercaptoethanol, EDTA, bromophenol blue
- Phosphatase inhibitors
- Did you heat the samples: temperature and time: 95oC for 5minutes, followed by 4oC-incubation for 3 mintues
4. Electrophoresis/Gel conditions/ Transfer conditions
- Reducing or non reducing gel: reducing
- Reducing agent: beta-mercaptoethanol
- Gel percentage : 8% running gel
- Transfer conditions: (Type of membrane, Protein transfer verified): PVDF, protein marker was completely transferred

5. Blocking conditions
- Buffer: TBST
- Blocking agent: milk, BSA, serum, what percentage: milk, 5%
- Incubation time: 1 hour
- Incubation temperature: room temperature
6. Primary Antibody
- Species:
- Reacts against:
- At what dilution(s) have you tested this antibody:
- What dilution buffer was used:
- Incubation time:
- Incubation temperature:
- What washing steps were done:
7. Secondary Antibody
- Species:Rabbit
- Reacts against: mouse and rat
- At what dilution(s) have you tested this antibody: 1/100 as suggested
- Incubation time: over 14 hours at 4oC
- Wash steps: membrane soaked in TBST and incubated in gentle shake for 5 minutes at room temperature; 3 times
- Fluorochrome or enzyme conjugate: enzyme conjugate; HRP
- Do you know whether the problems you are experiencing come from the secondary? Yes, and I see no problem with the secondary since it was applied on other antibody detection on the same membrane, and they all worked out, except anti-5HT2A
8. Detection method
ECl, ECl+, other detection method: ECL
9. Did you apply positive and negative controls along with the samples? Please specify.
I only got the internal control (beta actin) for quantitative analysis
10. Optimization attempts
- How many times have you tried the Western? 3
- Have you eliminated the possibility that any background bands could be due to the secondary antibody? (Run a “No primary” control): Yes, since there was no band found
- Do you obtain the same results every time e.g. are background bands always in the same place? The background always remained clear with no any band
- What steps have you altered?

PS the area framed by red box on the attached figure represents the 3 independent membranes for 5HT2A detection

Appreciate your assistance

Read More
Answer

Thank you for taking time to complete our questionnaire. I am sorry to hear that this antibody is not providing satisfactory results.

The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality.

Having reviewed this case, I would like to offer some suggestions to help optimize the results from ab66049 :

I would load slightly more sample on the gel, up to 30µg. This may help.
When testing our antibodies, our lab uses 5% BSA as a blocking reagent, so I recommend switching to this instead of milk. Some antibodies give stronger, more specific signals on blots blocked with BSA instead of milk, so doing this may improve the signal you are seeing. An example of the above is the western blot obtained with the Abcam GAPDH antibody ab9385 : https://www.abcam.com/ab9385.



Should the suggestions not improve the results, please do let me know.

In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within4 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund.

I hope this information is helpful, and I thank you for your cooperation.

Read More

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