Overview

  • Product name
  • Description
    Rabbit polyclonal to HCP1 / PCFT
  • Host species
    Rabbit
  • Specificity
    The immunogen used to raise this antibody has 80% homology with the human HCP1 / PCFT protein. Some customers have successfully used ab25134 in human tissue. Please contact Abcam Scientific Support for more information.
  • Tested applications
    Suitable for: IHC-P, ELISA, ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat
    Predicted to work with: Human
  • Immunogen

    Synthetic peptide corresponding to Mouse HCP1/ PCFT aa 400 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab25847)

  • Positive control
    • This antibody gave a positive signal in IF the following Formaldehyde fixed cell lines: NIH3T3. In Western Blot, this antibody gave a positive signal in NIH3T3 whole cell lysate and a positive signal in the following tissue lysates: Rat Liver; Rat Spleen; Rat Heart; Mouse Kidney.
  • General notes

    Previously labelled as HCP1.

Properties

Applications

Our Abpromise guarantee covers the use of ab25134 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-P 1/100. Perform heat mediated antigen retrieval via the microwave method before commencing with IHC staining protocol.
ELISA Use at an assay dependent concentration.
ICC/IF Use a concentration of 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 50 kDa (predicted molecular weight: 55 kDa).

Target

  • Function
    Has been shown to act both as an intestinal proton-coupled high-affinity folate transporter and as an intestinal heme transporter which mediates heme uptake from the gut lumen into duodenal epithelial cells. The iron is then released from heme and may be transported into the bloodstream. Dietary heme iron is an important nutritional source of iron. Shows a higher affinity for folate than heme.
  • Tissue specificity
    Expressed in kidney, liver, placenta, small intestine, spleen, retina and retinal pigment epithelium. Lower levels found in colon and testis. Very low levels in brain, lung, stomach, heart and muscle. In intestine, expressed in duodenum with lower levels in jejunum, ileum, cecum, rectum and segments of the colon.
  • Involvement in disease
    Defects in SLC46A1 are the cause of hereditary folate malabsorption (HFM) [MIM:229050]. HFM is a rare autosomal recessive disorder characterized by impaired intestinal folate absorption with folate deficiency resulting in anemia, hypoimmunoglobulinemia with recurrent infections, and recurrent or chronic diarrhea. In many patients, neurological abnormalities such as seizures or mental retardation become apparent during early childhood, attributed to impaired transport of folates into the central nervous system. When diagnosed early, the disorder can be treated by administration of folate. If untreated, it can be fatal and, if treatment is delayed, the neurological defects can become permanent.
  • Sequence similarities
    Belongs to the major facilitator superfamily. SLC46A family.
  • Cellular localization
    Apical cell membrane. Cytoplasm. Localizes to the apical membrane of intestinal cells in iron-deficient cells, while it resides in the cytoplasm in iron-replete cells.
  • Information by UniProt
  • Database links
  • Alternative names
    • G21 antibody
    • HCP 1 antibody
    • HCP1 antibody
    • Heme carrier protein 1 antibody
    • MGC9564 antibody
    • PCFT antibody
    • PCFT/HCP1 antibody
    • PCFT_HUMAN antibody
    • PDE7A antibody
    • Proton coupled folate transporter antibody
    • Proton-coupled folate transporter antibody
    • SLC46A1 antibody
    • Solute carrier family 46 (folate transporter) member 1 antibody
    • Solute carrier family 46 member 1 antibody
    see all

Images

  • All lanes : Anti-HCP1 / PCFT antibody (ab25134) at 1 µg/ml

    Lane 1 : Liver (Rat) Tissue Lysate
    Lane 2 : Spleen (Rat) Tissue Lysate
    Lane 3 : Heart (Rat) Tissue Lysate
    Lane 4 : Kidney (Mouse) Tissue Lysate
    Lane 5 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 55 kDa
    Observed band size: 50 kDa
    why is the actual band size different from the predicted?


    Exposure time: 8 minutes
  • All lanes : Anti-HCP1 / PCFT antibody (ab25134) at 1 µg/ml

    Lane 1 : Kidney (Mouse) Tissue Lysate
    Lane 2 : Kidney (Rat) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 55 kDa
    Observed band size: 50 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes
  • ICC/IF image of ab25134 stained NIH-3T3 cells. The cells were 4% paraformladhyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab25134, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
  • Image courtesy of Human Protein Atlas

    ab25134 staining HCP1 / PCFT in human stomach. Paraffin embedded human skin tissue was incubated with ab25134 (1/100 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.

    ab25134 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further results for this antibody can be found at www.proteinatlas.org

     

     

     

     

     

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References

This product has been referenced in:
  • Li H  et al. Physiological stress-induced corticosterone increases heme uptake via KLF4-HCP1 signaling pathway in hippocampus neurons. Sci Rep 7:5745 (2017). Read more (PubMed: 28720846) »
  • Takada T  et al. Selective accumulation of hematoporphyrin derivative in glioma through proton-coupled folate transporter SLC46A1. J Clin Biochem Nutr 54:26-30 (2014). IHC . Read more (PubMed: 24426187) »
See all 7 Publications for this product

Customer reviews and Q&As

1-10 of 14 Abreviews or Q&A

Answer


I have heard back from the lab with the following explanation regarding the membrane vs. cytoplasmic staining:
From what we can gather this protein can also show cytoplasmic localisation (swissprot).
It's very difficult to distinguish between cytoplasmic and membrane staining and we had very little data on this antibody as to the expected staining pattern. It seems to be consistent with other companies' data.
However, we welcome any feedback on the staining pattern from the customer for this antibody in IF.

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Answer

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 has caused. As requested, I have issued a free of charge replacement with the order number xxx with ab25134.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

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Question
Answer

Thank you for your reply.

A free of charge vial of ab98043 has been issued, which should arrive tomorrow.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

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Answer

DISCOUNT CODE: 100%000000

Expiration date: 14. October 2012


I am very pleased to hear you would like to accept our offer and test ab25134 in flow cytometry. This code will give you the value of ab25134 off a futuire order before the expiration date. To redeem this offer, please submit an Abreview for flow cytometryand include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active.
For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.


Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways:
1) Call to place your order and mention the code to our customer service department;
2) Include the code in your fax order;
3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

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Answer

Thank you very much for your interest in ab25134.

To our knowledge,ab25134 has not been tested in flow cytometry. Therefore, I can offer a discount off a future purchase if you buy ab25134 now, test it inflow cytometry and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of ab25134 off a future order.

If you are interested in this offer, please follow these steps:

1. Reply to this e-mail to let me know that you would like to proceed and test ab25134 in flow cytometry. I will then send a discount code. This code must be issued before purchasing ab25134 so please wait for my reply before ordering.

2. Purchase ab25134 either by phone, fax, or online (www.abcam.com).

3. Test it in flow cytometry.

4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any product ordered and the discount code is valid for 4 months after issue.
Please remember that submission of the Abreview is sufficient for the discount code to become active.

We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab25134 turns out to be unsuitable for flow cytometry, you will still receive the discount on your next purchase after your Abreview has been submitted.

Please let me know if you have any questions about this offer and I would be happy to help you further.

The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

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Answer

Thank you for contacting us.

I am sorry that this antibody did not perform as stated on the datasheet. I have asked our accounting department to issue a credit note for you, which can be redeemed against the invoice of a future order by passing it on to your purchasing department. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used. If you have questions on how to use the credit note, please contact our accounting department.

Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department.

The credit note ID is for your reference only and does not automatically guarantee the credit.

I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

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Answer

Thank you for following up with your results.

I am sorry that the blocking peptide experiments did not demonstrate the specificity of the bands you were detecting in your samples. I have checked our available stock and we only have the lot you currently have available. I would be happy to offer you another vial from this lot or a credit. It seems that the other antibodies to this target have not been validated for WB and mouse.

I look forward to your reply so that I may assist you further. Please do not hesitate to contact me if you have any additional questions.

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Answer

Thank you for your reply.


The new results you obtained are quite interesting. The image on our website is not representing data from the current lot, but is representative of the expected results with this antibody.


Per your request, I have sent you the HCP1 peptide ab25847 free of charge. This should arrive to you early next week.


I hope this will be helpful for your experiments. Please do not hesitate to contact me if you have any additional questions.

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Answer

Thank you for contacting Abcam regarding ab25134.


I have asked the lab to retest this antibody. We observed an ˜60 kDa in most Hu tissues and an ˜50 kDa in Ms and Rat tissues, consistent with our ds images. We used 5% BSA to block so it's possible if you are using milk you may see a different WB profile.


If you would provide an order number I would be happy to offer a replacement or credit. Please let me know which you would prefer.


I look forward to your reply so that I may assist you further. Please do not hesitate to contact us if you have any additional questions.

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Answer

We have previously been in contact regarding ab25134, which was not providing satisfactory results.

I would appreciate if you could let me know whether satisfactory results were obtained with the antibody.

I wish you the best of luck with your research and please do not hesitate to contact us if you have any further questions.

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1-10 of 14 Abreviews or Q&A

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