Overview

  • Product name
  • Description
    Guinea pig polyclonal to Substance P
  • Host species
    Guinea pig
  • Tested applications
    Suitable for: IHC-FoFr, ICC/IF, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide:

    CRPKPQQFFGLM

    , corresponding to amino acids 1-11 of Rat Substance P (Peptide available as ab38217.)

  • General notes


    This gene encodes four products of the tachykinin peptide hormone family, substance P and neurokinin A, as well as the related peptides, neuropeptide K and neuropeptide gamma. These hormones are thought to function as neurotransmitters which interact with nerve receptors and smooth muscle cells. They are known to induce behavioral responses and function as vasodilators and secretagogues. Alternative splicing of exons 4 and/or 6 produces four known products of undetermined significance.

Properties

Associated products

Applications

Our Abpromise guarantee covers the use of ab10353 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 1/1000.
ICC/IF Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.

Target

  • Function
    Tachykinins are active peptides which excite neurons, evoke behavioral responses, are potent vasodilators and secretagogues, and contract (directly or indirectly) many smooth muscles.
  • Sequence similarities
    Belongs to the tachykinin family.
  • Cellular localization
    Secreted.
  • Information by UniProt
  • Database links
  • Alternative names
    • C-terminal-flanking peptide antibody
    • Hs.2563 antibody
    • neurokinin 1 antibody
    • neurokinin 2 antibody
    • neurokinin A antibody
    • neurokinin alpha antibody
    • Neuromedin L antibody
    • neuropeptide gamma antibody
    • neuropeptide K antibody
    • NK2 antibody
    • NKA antibody
    • NKNA antibody
    • NPK antibody
    • PPT antibody
    • preprotachykinin antibody
    • protachykinin antibody
    • protachykinin-1 antibody
    • Substance K antibody
    • SubstanceP antibody
    • TAC1 antibody
    • TAC2 antibody
    • TAC2, formerly antibody
    • Tachykinin 1 antibody
    • tachykinin 2 antibody
    • tachykinin 2, formerly antibody
    • Tachykinin precursor 1 antibody
    • tachykinin, precursor 1 (substance K, substance P, neurokinin 1, neurokinin 2, neuromedin L, neurokinin alpha, neuropeptide K, neuropeptide gamma) antibody
    • Tachykinin1 antibody
    • TKN1_HUMAN antibody
    see all

Images

  • ab10353 staining Substance P in Human brain stem tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 1% BSA for 10 minutes at 21°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/200 in blocking buffer) for 2 hours at 22°C. A Biotin-conjugated Rabbit anti-guinea pig polyclonal (1/200) was used as the secondary antibody.

    See Abreview

  • Immuofluorescent detection of Substance P in rat spinal cord dorsal horn (red fluorescence). DAPI (blue) was used as counter stain.
  • Immunohistochemical analysis of rat paw skin tissue, staining Substance P with ab10353 at 1/1000 dilution.
  • ab10353 staining Substance P in Mouse brain stem  tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 1% BSA for 10 minutes at 21°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/400 in blocking buffer) for 2 hours at 21°C. A Biotin-conjugated Rabbit anti-Guinea pig polyclonal (1/200) was used as the secondary antibody.

    See Abreview

  • ab10353 staining Substance P in mouse spinal cord (red fluorescence) by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue was counterstained with Fluoro Nissl Green (green fluorescence).
  • Sustance P staining of trigeminal ganglia from P3-P5 rat pups.

References

This product has been referenced in:
  • Nagy JI  et al. Connexin36 Expression in Primary Afferent Neurons in Relation to the Axon Reflex and Modality Coding of Somatic Sensation. Neuroscience 383:216-234 (2018). Read more (PubMed: 29746988) »
  • Bright FM  et al. Normative distribution of substance P and its tachykinin neurokinin-1 receptor in the medullary serotonergic network of the human infant during postnatal development. Brain Res Bull 137:319-328 (2018). IF . Read more (PubMed: 29331576) »
See all 19 Publications for this product

Customer reviews and Q&As

1-10 of 10 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (colon)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mm sodium citrate PH 6
Permeabilization
No
Specification
colon
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Apr 28 2016

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Sample
Goat Tissue sections (Small intestine)
Specification
Small intestine
Permeabilization
No
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Nov 19 2014

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1%
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Sample
Pig Tissue sections (Colon)
Specification
Colon
Permeabilization
No
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Nov 18 2014

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: c
Sample
Dog Tissue sections (Colon)
Specification
Colon
Permeabilization
No
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Nov 18 2014

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Sample
Marmoset (common) Tissue sections (Hippocampus)
Specification
Hippocampus
Permeabilization
No
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Nov 03 2014

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Sample
Human Tissue sections (Brainstem)
Specification
Brainstem
Permeabilization
No
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Nov 03 2014

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric aicd
Sample
Cat Tissue sections (Spinal cord)
Specification
Spinal cord
Permeabilization
No
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Nov 03 2014

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Sample
Rat Tissue sections (Brain)
Specification
Brain
Permeabilization
No
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Nov 03 2014

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Sample
Mouse Tissue sections (Brainstem)
Specification
Brainstem
Permeabilization
No
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Nov 03 2014

Question

LOT NUMBER GR29977-3 ORDER NUMBER 998323 DESCRIPTION OF THE PROBLEM Non-specific staining SAMPLE Drd2-EGFP transgenic mice PRIMARY ANTIBODY Concentration or dilution 1:50~200 Diluent buffer same as blocking solution Incubation time 24hrs or 48hrs at RT Wash Buffer 0.5%normal horse serum in 0.5%TBST(triton-100) Number of washes 5min x 4times DETECTION METHOD confocal (lsm700) imaging POSITIVE AND NEGATIVE CONTROLS USED Positive control dorsal striatum Negative control – without primary antibody(didn’t show the signal) ANTIBODY STORAGE CONDITIONS -20°C FIXATION OF SAMPLE Fixation step Yes/No yes If yes: Fixative agent and concentration 4% paraformylaldehyde Fixation time 4hrs of post fixation and 30% sucrose in PBS(FOR CRYOSTAT) Fixation temperature 4°C ANTIGEN RETRIEVAL endogenous quenching(0.3%H2O2, 50%Methanol), Citrate Buffer (10mM Citric Acid, 0.05% Tween 20, pH 6.0) PERMEABILIZATION STEP - BLOCKING CONDITIONS 10% normal horse serum in 0.5% TBST(TritonX-100) Blocking time 30min Blocking temperature RT Endogenous peroxidases blocked? YES Endogenous biotins blocked? NO SECONDARY ANTIBODY Bethyl A60-110D3 Species: GOAT Isotype:IgG Reacts against: guinea PIG Concentration or dilution 1:400 Diluent buffer same as blocking solution Incubation time 1hr at RT Fluorochrome or enzyme conjugate DyLight550 Wash Buffer 0.5%normal horse serum in 0.5%TBST(triton-100) Number of washes 5min x 4times HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 5 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? I have altered blocking concentration and primary incubation time and temperature. ADDITIONAL NOTES I performed immunohistochemistry with substance P antibody to Dopamine 2receptor eGFP transgenic mice. In order to substance P is not involved dopamine indirect pathway it should not be localized to D2R eGFP mice. But these experiments have shown that substance P was detected on D2R eGFP specific neurons. So I think the specificity of this antibody worked wrongly. For this reason, I want to replace this to other product.

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Answer

Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody. The details you have kindly provided will provide us with vital information for our monitoring of product quality. I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful. Reviewing the details, I am sorry there are no further tips to provide on this occasion to help improve the results. I can suggest you may regrettably have received a bad vial. I apologize for the inconvenience and am pleased to offer you a free of charge replacement or credit note in compensation. Although there are no major tips to provide, I would like to recommend to consider the following for future experiments, which I hope will help: 1. The datasheet suggests a dilution of 1:1000 for this application. I can recommend to try this lower concentration in order to reduce any non specific staining. 2. I can recommend to try different time points for antigen retrieval if this has not already been tried. The antigen retrieval step can sometimes require optimization for best results. 3. Are you aware of any publications to confirm that D2R is never colocalised with substance P? Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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