Overview

  • Product name

    Anti-RFC antibody
  • Description

    Rabbit polyclonal to RFC
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ELISA, WB, IHC-P, ICC/IFmore details
  • Species reactivity

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

    Synthetic peptide corresponding to Human RFC aa 1-50 (N terminal).
    Sequence:

    MVPSSPAVEKQVPVEPGPDPELRSWRHLVCYLCFYGFMAQIRPGESFITP

  • Positive control

    • Jurkat cell lysate. Human kidney.
  • General notes

    Protein previously labeled as SLC19A1.

Properties

Applications

Our Abpromise guarantee covers the use of ab62302 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
ELISA Use at an assay dependent concentration.
WB Use a concentration of 5 µg/ml. Detects a band of approximately 66 kDa (predicted molecular weight: 66 kDa). Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.
IHC-P Use a concentration of 4 - 8 µg/ml.
ICC/IF Use a concentration of 5 µg/ml.

Target

  • Function

    Transporter for the intake of folate. Uptake of folate in human placental choriocarcinoma cells occurs by a novel mechanism called potocytosis which functionally couples three components, namely the folate receptor, the folate transporter, and a V-type H(+)-pump.
  • Tissue specificity

    Placenta, liver, and to a much smaller extent, in lung.
  • Sequence similarities

    Belongs to the reduced folate carrier (RFC) transporter (TC 2.A.48) family.
  • Post-translational
    modifications

    Heavily glycosylated.
  • Cellular localization

    Membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • CHMD antibody
    • FLOT 1 antibody
    • FLOT1 antibody
    • Folate transporter 1 antibody
    • FOLT antibody
    • IFC 1 antibody
    • IFC-1 antibody
    • IFC1 antibody
    • Intestinal folate carrier 1 antibody
    • Intestinal folate carrier antibody
    • OTTHUMP00000115459 antibody
    • OTTHUMP00000115460 antibody
    • Placental folate transporter antibody
    • Reduced folate carrier antibody
    • Reduced folate carrier protein antibody
    • REFC antibody
    • RFC 1 antibody
    • RFC antibody
    • RFC1 antibody
    • S19A1_HUMAN antibody
    • SLC19A1 antibody
    • Solute carrier family 19 member 1 antibody
    see all

Images

  • RFC antibody (ab62302) at 5.0µg/ml (in 5% skim milk / PBS buffer) + Jurkat cell lysate at 10 µg

    Secondary
    HRP conjugated anti-Rabbit IgG at 1/50000 dilution

    Predicted band size: 66 kDa
    Observed band size: 66 kDa



    Gel concentration: 8%
  • ICC/IF image of ab62302 stained MCF7 cells. The cells were 4% PFA fixed (10mins) 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 (ab62302, 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labelling RFC with ab62302 at 4-8µg/ml. Arrows indicate positively stained epithelial cells of the renal tubule. Magnification: 400X.

References

This product has been referenced in:

See all 6 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Abreviews
Application
Western blot
Sample
Rat Tissue lysate - other (Brain)
Gel Running Conditions
Reduced Denaturing (12% gel)
Loading amount
30 µg
Specification
Brain
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5µg/mL · Temperature: 25°C

Aniv Mann

Verified customer

Submitted Oct 30 2015

Application
Western blot
Sample
Human Cell lysate - whole cell (THLE2 liver cell line)
Gel Running Conditions
Reduced Denaturing (Polyacrylamide gel precast 12-4%)
Loading amount
20 µg
Specification
THLE2 liver cell line
Blocking step
Li-cor Blocking Buffer as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jul 04 2014

Answer

Thank you for your message,

I apologize for any confusion. We do sell ab62302 Anti-SLC19A1 antibodyin lyophillised form now. I can suggest in this case youhave receivedan older vial which has been reconstituted already.This can be used as normal.

I hope this will be helpful. If you have any further questions or concerns, please do not hesitate to contact me.

Read More

Question

Please find below the details of customer's complaint. Attachedisthe image.Please advise.
Antibody code: ab62302 SLC19A1



Batch number:


Antibody storage conditions (temperature/reconstitution etc) stock at – 20 0 c.

Description of the problem (high background, wrong band size, more bands, no band etc.) more bands, each replicate looks different (non consistent result)

Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.)BEWO cells Hu /whole cell lysate

Sample preparation (Buffer/Protease inhibitors/Heating sample etc.)lysis buffer V(1 M Tris·HCl, pH7.5 10%SDS 5mg/ml DNase I, 1 M MgCl2, 50 mg/ml PMSF, and protease inhibitor cocktail). Sample buffer –laemmli buffer

Not heated!

Amount of protein loaded 30ug total protein/lane

Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.)10% reducing gel

Transfer and blocking conditions (Buffer/time period, Blocking agent etc.)Wet transfer 1.5h ,blocking with 5% skim milk/TBST -0.1% For 1h

Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step) abcam Rabbit polyclonal reacts with Hu, diluted Ab 1:200 in 5%BSA/TBS –T 0.1%, incubation ON 40c . 3X washes with TBST 10' each wash

Also 1:100 and 1:500 were tried and the same result was observed.

Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step) Jackson ImmunoReserch,peroxidase conjugate goat anti rabbit IgG (H+L) diluted 1:20000 5% skim milk/TBS-T0.1%



1:10000, 1:15000, 1:20,000 dilutions of the secondary were used.

Detection method (ECL, ECLPlus etc.) ECL sigma luminol +coumaric acid +H2o2/tris pH 7.5

Positive and negative controls used (please specify ) not used



Optimization attempts (problem solving)



How many times have you tried the Western? 5 times

Have you run a "No Primary" control? yes

Do you obtain the same results every time? e.g. are the background bands always in the same place? yes

What steps have you altered?different dilution of 1st 2nd Ab


Thanks in advance for your assistance.

Read More
Answer

Thank you and your customer for taking the time to complete our questionnaire and contact us. I am sorry to hear your customer has had difficulty obtaining satisfactory results from this antibody.
The details your customer has kindly provided will enable us to investigate this case for your customer and this is also helpful in our records for monitoring of quality.
Reviewing this case, I would like to offer some suggestions to help optimise the results from ab62302.
1.) I would like to suggest to decrease the boiling temperature to 65C and to prolong the boiling time up to 10 min. Your target is a multi-pass membrane protein, and these proteins have the unfortunate tendency to aggregate during boiling periods with high temperatures and beta- mercaptoethanol due to their hydrophobic nature.
2.) I also recommend to use a positive control. For example jurkat cells, they showed on clean band in our test experiments as shown on the datasheet.
3.) I can suggest not to mix blocking agent but to either use milk or BSA.
4.) I would like to suggest to use less SDS. 10% seems very high. What is the reason that this much SDS is used? Standart lysis buffer contains ca 0,1% SDS. Also DNase should not be working at SDS concentrations this high. There are Nucleases available that are SDS resistant, but I am unsure if they can withstand 10% SDS.
5.) I also recommend a no primary control tom exclude the possibility that the extra bands are due to the secondary antibody cross reacting un-specifically.
6.) This protein can be glycosylated and phosphorylated and the multiple bands could be different states of phophorylatipn and/or glycosylation of the protein that are independent of the treatment.
I would also appreciate if your customer can confirm some further details:
1.) Could your customer please provide an image of the whole blot?
2.) What was the treatment and what is the expected result of the treatment?
We are happy to offer this technical support. In the event that a product is not functioning in the applications cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.
I hope this information is helpful, thank you and your customer for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.

Read More
Application
Western blot
Sample
Rat Cell lysate - whole cell (choroidal epithelial Z310 cells)
Gel Running Conditions
Reduced Non-Denaturing (Native) (gel 10%)
Loading amount
50 µg
Specification
choroidal epithelial Z310 cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted May 12 2011

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