Overview

  • Product name

    Anti-Folate Binding Protein/FBP antibody
    See all Folate Binding Protein/FBP primary antibodies
  • Description

    Rabbit polyclonal to Folate Binding Protein/FBP
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, WB, IHC-Pmore details
  • Species reactivity

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

    Synthetic peptide corresponding to Human Folate Binding Protein/FBP aa 50-150 conjugated to keyhole limpet haemocyanin.
    Database link: P15328
    (Peptide available as ab87124)

  • Positive control

    • This antibody gave a positive signal in Mouse Cerebellum Tissue Lysate. This antibody gave a positive result in IF in the following Formaldehyde fixed cell line: HepG2.
  • General notes

    Previously labelled as Folate Binding Protein

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab67422 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
ICC/IF Use a concentration of 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 28 kDa (predicted molecular weight: 30 kDa). Please note, we have been advised by some customers that ab67422 is unable to detect the human version of this protein in western blot. Please contact our scientific support services if you have any queries regarding this antibody.
IHC-P Use a concentration of 10 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Target

  • Function

    Binds to folate and reduced folic acid derivatives and mediates delivery of 5-methyltetrahydrofolate to the interior of cells.
  • Tissue specificity

    Exclusively expressed in tissues of epithelial origin. Expression is increased in malignant tissues. Expressed in kidney, lung and cerebellum.
  • Involvement in disease

    Defects in FOLR1 are the cause of neurodegeneration due to cerebral folate transport deficiency (NCFTD) [MIM:613068]. NCFTD is an autosomal recessive disorder resulting from brain-specific folate deficiency early in life. Onset is apparent in late infancy with severe developmental regression, movement disturbances, epilepsy, and leukodystrophy. Note=Recognition and diagnosis of this disorder is critical because folinic acid therapy can reverse the clinical symptoms and improve brain abnormalities and function.
  • Sequence similarities

    Belongs to the folate receptor family.
  • Post-translational
    modifications

    Eight disulfide bonds are present.
    The secreted form is derived from the membrane-bound form either by cleavage of the GPI anchor, or/and by proteolysis catalyzed by a metalloprotease.
  • Cellular localization

    Cell membrane. Secreted.
  • Information by UniProt
  • Database links

  • Alternative names

    • adult antibody
    • Adult folate binding protein antibody
    • Adult folate-binding protein antibody
    • FBP antibody
    • Folate Binding Protein antibody
    • Folate Receptor 1 Adult antibody
    • Folate receptor 1 antibody
    • Folate Receptor 1 Precursor antibody
    • Folate receptor adult antibody
    • Folate receptor alpha antibody
    • Folate receptor antibody
    • FOLR antibody
    • FOLR1 antibody
    • FOLR1_HUMAN antibody
    • FR alpha antibody
    • FR-alpha antibody
    • FRalpha antibody
    • KB cells FBP antibody
    • MOV18 antibody
    • Ovarian cancer associated antigen antibody
    • Ovarian tumor associated antigen antibody
    • Ovarian tumor associated antigen MOv18 antibody
    • Ovarian tumor-associated antigen MOv18 antibody
    see all

Images

  • Anti-Folate Binding Protein/FBP antibody (ab67422) at 1/1 dilution + Cerebellum Mouse Tissue Lysate at 10 µg

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 30 kDa
    Observed band size: 28 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 16 kDa (possible cleavage fragment), 22 kDa (possible cleavage fragment)



    We hypothesize that the 28, 22 and 16 kDa bands represent the propeptide with signal sequence, propeptide without signal sequence and mature protein, respectively.
  • IHC image of Folate Binding Protein/FBP staining in human kidney carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab67422, 10µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

  • ab67422 stained HepG2 cells. The cells were 4% formaldehyde 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 ab67422 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 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.

References

This product has been referenced in:

  • Martin JB  et al. Folic acid modifies the shape of epithelial cells during morphogenesis via a Folr1 and MLCK dependent mechanism. Biol Open 8:N/A (2019). Read more (PubMed: 30670450) »
  • Siwowska K  et al. Folate Receptor-Positive Gynecological Cancer Cells: In Vitro and In Vivo Characterization. Pharmaceuticals (Basel) 10:N/A (2017). Read more (PubMed: 28809784) »
See all 4 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Question

Dear Tech Support Team Please find below the details of customer's complaint. Attached is the image. Please advise.
Antibody storage conditions (temperature/reconstitution etc) at – 20 0 c.

Description of the problem (high background, wrong band size, get the cleavage fragment

The band of approximately 28 kDa is not observed at all.

Sample BEWO cells Hu ,whole cell lysate

Sample preparationLysis buffer 0.01 M Tris·HCl, pH7.5 0.1%SDS 0.05mg/ml DNase I,0.0 1 M MgCl2,0. 5 mg/ml PMSF, and protease inhibitor cocktail). Sample buffer –laemmli buffer

Samples Not heated!

Amount of protein loaded 30ug total protein/lane

Electrophoresis/Gel conditions - 10% reducing gel

Transfer and blocking conditions Wet transfer 1.5h ,blocking with 5% skim milk/TBST -0.1% For 1h

Primary Antibody abcam, Rabbit polyclonal reacts with Hu and Ms, diluted Ab 1:1000 1ug/ml in 5%BSA/TBS –T 0.1%, incubation O/N 40c . 3X washes with TBST 10' each wash

Secondary Jackson ImmunoReserch ,peroxidase conjugate goat anti rabbit IgG (H+L) diluted 1:10000 5% skim milk/TBS-T0.1%

Detection method ECL sigma luminol +coumaric acid +H2o2/tris pH 7.5

Positive and negative controls used (please specify ) not use



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 oft 2nd Ab 1:15000, 1:5000
Thanks in advance for your assistance and reply.

Read More
Answer

Thank you for contacting us and for sending the details of the customer's complaint.
I am sorry that this antibody has not given the expected results. We have actually received reports that this antibody does not work well in Western blot with human lysates, and we are investigating the issue. Due to these concerns, I am happy to offer a replacement to the customer or a credit or refund if they prefer. We also have ab125030 which is guaranteed to work in Western blot with human samples-
https://www.abcam.com/Folate-Binding-Protein-antibody-EPR4708-2-ab125030.html
Please let me know how your customer would like to proceed and I will process it immediately. I look forward to hearing from you. If you have any questions or if there is anything else that we can do for you, please let me know.

Read More
Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (Lung cancer)
Permeabilization
No
Specification
Lung cancer
Fixative
Acetone

Han-Byul Lee

Verified customer

Submitted May 28 2019

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Kidney (untreated CD1 mouse))
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate Buffer pH6, 20 min, 98C
Permeabilization
No
Specification
Kidney (untreated CD1 mouse)
Blocking step
Dako S2023 as blocking agent for 10 minute(s) · Concentration: 0µg/mL · Temperature: RT°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Oct 30 2015

Question

Here are the answers:
Antibody code: ab67422
Batch number: XXXX
Antibody storage conditions (temperature/reconstitution etc) stock at – 20 0 c.
Description of the problem (wrong bands size,
Sample Ms liver extract
Sample preparation Lysis buffer 10m M Tris·HCl, pH7.5 250mM sucrose ,10mM KH2PO4, 5mM EDTA, PMSF, and protease inhibitor cocktail
Samples were heated!
Amount of protein loaded 30ug total protein/lane
Electrophoresis/Gel conditions - 10% reducing gel
Transfer and blocking conditions Wet transfer 1.5h ,blocking with 5% skim milk/TBST -0.1% For 1h
Primary Antibody abcam, Rabbit polyclonal reacts with Hu and Ms, diluted Ab 1:1000 1ug/ml in 5%BSA/TBS –T 0.1%, incubation O/N 40c . 3X washes with TBST 10' each wash
Secondary Jackson ImmunoReserch ,peroxidase conjugate goat anti rabbit IgG (H+L) diluted 1:10000 5% skim milk/TBS-T0.1%
Detection method ECL sigma luminol +coumaric acid +H2o2/tris pH 7.5
Positive and negative controls used (please specify ) not use
Optimization attempts (problem solving)
How many times have you tried the Western? 2 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?
Additional Notes The customer performed WB with mouse liver lysates . According to the datasheet this antibody should detect a band of approximately 28 kDa, but the customer observes 2 bands: 60kDa and 180 kDa. Kindly advise the meaning of this result and how it can be explained. The image of the gel is attached. Thank you for the assistance.

Read More
Answer

Thank you for taking the time to complete our questionnaire and sending the images. I am sorry to hear you have had difficulty obtaining satisfactory results from our products.
Reviewing this case, I would like to offer some suggestions to help optimise the results from ab67422:
-This protein is exclusively expressed in tissues of epithelial origin. I have checked its expression in liver tissue and confirmed it is not expressed in it:
http://www.ncbi.nlm.nih.gov/UniGene/ESTProfileViewer.cgi?uglist=Mm.2135
Do you have any evidence, references or publications confirming the presence of folate binding protein?
In order to check the antibody’s performance I would suggest running a positive control in parallel, like mouse kidney tissue lysate, or Mouse Cerebellum Tissue Lysate.
-Please make sure the denaturing and reducing conditions are optimised for this particular case. As a lysis buffer, we usually recommend RIPA buffer which has a high denaturing level.
-I would also recommend decreasing the antibodies concentrations, to 1/500 dilution for the primary and 1/5000 for the secondary.
- Have you checked neither of the unspecific bands corresponds to the secondary?
I would much appreciate if you could please provide the order number.
I hope this suggestions help improve the results from the antibody, if they didn’t our Abpromise would cover it, and a free of charge replacement or credit note will be offered.

Read More

Answer

Thank you for contacting us and sending the image.
Without further details it is very difficult to say where these bands observed come from. I am very happy to help you, and for that, I would appreciate if you could please send me more protocol details. I send you a questionnaire that only take about 5 minutes to be completed, and could help us understand the results obtained.
Once we receive the completed questionnaire, we will look at the protocol and see if there are any suggestions we can make that may improve the results. If no suggestions can be made to the protocol, and the antibody was purchased within the last 6 months, it is covered by our Abpromise guarantee, and therefore I can offer you a free of charge replacement or a credit note for it.
I look forward to receiving your reply. Please do not hesitate to contact us if you need any more advice or information.

Read More

Answer

Thank you for your enquiry. I have also run a BLAST homology and the results is as below. gi|148225128|ref|NP_001086114.1| folate receptor 1 (adult) [Xenopus laevis] gi|49257600|gb|AAH74206.1| MGC82130 protein [Xenopus laevis] Length=243 GENE ID: 444543 folr1 | folate receptor 1 (adult) [Xenopus laevis] (10 or fewer PubMed links) Score = 32.3 bits (72), Expect = 0.35, Method: Compositional matrix adjust. Identities = 12/16 (75%), Positives = 14/16 (88%), Gaps = 0/16 (0%) It seems that the percentage of homology is 75%. We can usually expect positive reactivity if the homology is more than 90% but having a low homology percentage does not necessary mean no cross reactivity whatsoever. This is simply a way to check for suitability. I hope the information above will be helpful. If there is anything else that I can help you with please do not hesitate to contact me. Have a nice day!

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