Overview

  • Product name

  • Description

    Rabbit polyclonal to HV1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ELISA, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Synthetic peptide corresponding to Human HV1 (C terminal) conjugated to keyhole limpet haemocyanin. (NP_115745.2).
    (Peptide available as ab214795)

  • Positive control

    • Human spleen and thymus tissues.
  • General notes

    Protein previously labeled as HVCN1.

Properties

Applications

Our Abpromise guarantee covers the use of ab117520 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 Use a concentration of 0.5 - 1 µg/ml. Predicted molecular weight: 32 kDa.Can be blocked with HV1 peptide (ab214795).
ELISA Use at an assay dependent concentration.
IHC-P Use a concentration of 5 µg/ml.

Target

  • Function

    Mediates the voltage-dependent proton permeability of excitable membranes. Forms a proton-selective channel through which protons may pass in accordance with their electrochemical gradient. Proton efflux, accompanied by membrane depolarization, facilitates acute production of reactive oxygen species in phagocytosis.
  • Tissue specificity

    Enriched in immune tissues, such as lymph nodes, B-lymphocytes, monocytes and spleen.
  • Sequence similarities

    Belongs to the hydrogen channel family.
  • Domain

    The segment S4 is probably the voltage-sensor and is characterized by a series of positively charged amino acids at every third position. Unlike other voltage-gated ion channels it lacks the pore domain.
    The C-terminal coiled coil region mediates homodimerization and cooperative channel gating. It is essential for normal subcellular localization.
  • Post-translational
    modifications

    Phosphorylation may enhance channel gating.
  • Cellular localization

    Membrane. Cell membrane. Detected mainly at intracellular membranes upon overexpression in HeLa cells (PuMed:20147290), but not in other cell types.
  • Information by UniProt
  • Database links

  • Alternative names

    • HV 1 antibody
    • HV1 antibody
    • HVCN 1 antibody
    • HVCN1 antibody
    • HVCN1_HUMAN antibody
    • Hydrogen voltage gated channel 1 antibody
    • Hydrogen voltage-gated channel 1 antibody
    • Voltage gated hydrogen channel 1 antibody
    • Voltage sensor domain only protein antibody
    • Voltage sensor domain-only protein antibody
    • Voltage-gated hydrogen channel 1 antibody
    • VSOP antibody
    see all

Images

  • ab117520, at 5 µg/ml, staining HV1 in Formalin-fixed, Paraffin-embedded Human spleen tissue by Immunohistochemistry.

  • ab117520, at 5 µg/ml, staining HV1 in Formalin-fixed, Paraffin-embedded Human thymus tissue by Immunohistochemistry.

References

This product has been referenced in:

  • Patel B  et al. Voltage gated proton channels modulate mitochondrial reactive oxygen species production by complex I in renal medullary thick ascending limb. Redox Biol N/A:101191 (2019). Read more (PubMed: 31060879) »
See 1 Publication for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Answer

I am sorry for any confusion.

This protocol is provided from the lab which supplies ab117520 directly to us. It is the same protocol they use to test all their antibodies in IHC butrest assured it is not arandom protocol from IHC world.

Perhaps it would be helpful if you could provide your IHC protocol. I can pass it along to our supplying lab and get their opinon on any potential changes that may improve your results.

Thanks in advance for the additional information.

Read More

Answer

Thank you for your enquiry. Below please find the IHC protocol requested.

I hope this is helpful. Please contact me again if you have any further questions.

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Tissue Preparation

Formalin fixation and embedding in paraffin wax



Tissue Sectioning

Make 4-µm sections and place on pre-cleaned and charged microscope slides.
Heat in a tissue-drying oven for 45 minutes at 60°C



Deparaffinization

Wash slides in 3 changes of xylene – 5 minutes each at room temperature.



Rehydration

Wash slides in 3 changes of 100% alcohol – 3 minutes each at room temperature.
Wash slides in 2 changes of 95% alcohol – 3 minutes each at room temperature.
Wash slides in 1 change of 80% alcohol – 3 minutes at room temperature.
Rinse slides in gentle running distilled water – 5 minutes at room temperature.



Antigen retrieval

Steam slides in 0.01 M sodium citrate buffer, pH 6.0 at 99-100°C - 20 minutes
Remove from heat and let stand at room temperature in buffer - 20 minutes
Rinse in 1X TBS with Tween (TBST) – 1 minute at room temperature.



Immunostaining

Do not allow tissues to dry at any time during the staining procedure.
Apply a universal protein block – 20 minutes at room temperature.
Drain protein block from slides, apply diluted primary antibody – 45 minutes at room temperature.
Rinse slides in 1X TBST - 1 minute at room temperature.
Apply a biotinylated secondary antibody (specific to the host of the primary antibody) - 30 minutes at room temperature.
Rinse slides 1X TBST – 1 minute at room temperature.
Apply alkaline phosphatase streptavidin – 30 minutes at room temperature.
Rinse slides in 1X TBST - 1 minute at room temperature.
Apply alkaline phosphatase chromogen substrate – 30 minutes at room temperature.
Wash slides in distilled water – 1 minute at room temperature.



Dehydrate

This method should only be used if the chromogen substrate is alcohol insoluble.
Wash slides in 2 changes of 80% alcohol – 1 minute each at room temperature.
Wash slides in 2 changes of 95% alcohol – 1 minute each at room temperature.
Wash slides in 3 changes of 100% alcohol – 1 minute each at room temperature.
Wash slides in 3 changes of xylene – 1 minute each at room temperature.
Apply coverslip

Read More

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