Anti-Folate Binding Protein/FBP antibody (ab67422)
Key features and details
- Rabbit polyclonal to Folate Binding Protein/FBP
- Suitable for: ICC/IF, WB, IHC-P
- Reacts with: Mouse, Human
- Isotype: IgG
Overview
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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
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Immunogen
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Positive control
- WB: Mouse cerebellum tissue lysate, JAR and HeLa cell lysates. ICC/IF: HepG2 cells.
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General notes
Previously labelled as Folate Binding Protein
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Properties
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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
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Isotype control
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KO cell lines
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KO cell lysates
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KO cell pellets
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
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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
modificationsEight 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
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Database links
- Entrez Gene: 2348 Human
- Entrez Gene: 171049 Rat
- Omim: 136430 Human
- SwissProt: P15328 Human
- SwissProt: P35846 Mouse
- Unigene: 73769 Human
- Unigene: 2135 Mouse
- Unigene: 6912 Rat
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Alternative names
- adult antibody
- Adult folate binding protein antibody
- Adult folate-binding protein antibody
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Folate Binding Protein/FBP antibody (ab67422)
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.
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Western blot - Anti-Folate Binding Protein/FBP antibody (ab67422)All lanes : Anti-Folate Binding Protein/FBP antibody (ab67422) at 1/500 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : FOLR1 knockout HeLa cell lysate
Lane 3 : JAR cell lysate
Lane 4 : MDA-MB-231 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 30 kDa
Observed band size: 38 kDa why is the actual band size different from the predicted?Lanes 1-4: Merged signal (red and green). Green - ab67422 observed at 38 kDa. Red - loading control, ab7291 observed at 52 kDa.
ab67422 Anti-Folate Binding Protein/FBP antibody was shown to specifically react with Folate Binding Protein/FBP in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264921 (knockout cell lysate ab257270) was used. Wild-type and Folate Binding Protein/FBP knockout samples were subjected to SDS-PAGE. ab67422 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
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Western blot - Anti-Folate Binding Protein/FBP antibody (ab67422)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. -
Immunocytochemistry/ Immunofluorescence - Anti-Folate Binding Protein/FBP antibody (ab67422)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.
Protocols
Datasheets and documents
References (6)
ab67422 has been referenced in 6 publications.
- 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). PubMed: 30670450
- Alam C et al. Upregulation of reduced folate carrier by vitamin D enhances brain folate uptake in mice lacking folate receptor alpha. Proc Natl Acad Sci U S A 116:17531-17540 (2019). PubMed: 31405972
- Siwowska K et al. Folate Receptor-Positive Gynecological Cancer Cells: In Vitro and In Vivo Characterization. Pharmaceuticals (Basel) 10:N/A (2017). PubMed: 28809784
- Meredith M et al. Growing Mouse Oocytes Transiently Activate Folate Transport via Folate Receptors As They Approach Full Size. Biol Reprod 94:125 (2016). PubMed: 27122634
- Ocak M et al. Folate receptor-targeted multimodality imaging of ovarian cancer in a novel syngeneic mouse model. Mol Pharm 12:542-53 (2015). PubMed: 25536192
- Castiglioni V et al. Immunohistochemical Characterization of a Renal Nephroblastoma in a Trp53-mutant and Prolyl Isomerase 1-deficient Mouse. J Toxicol Pathol 26:423-7 (2013). PubMed: 24526816
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Folate Binding Protein/FBP antibody (ab67422)
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.
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Western blot - Anti-Folate Binding Protein/FBP antibody (ab67422)All lanes : Anti-Folate Binding Protein/FBP antibody (ab67422) at 1/500 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : FOLR1 knockout HeLa cell lysate
Lane 3 : JAR cell lysate
Lane 4 : MDA-MB-231 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 30 kDa
Observed band size: 38 kDa why is the actual band size different from the predicted?Lanes 1-4: Merged signal (red and green). Green - ab67422 observed at 38 kDa. Red - loading control, ab7291 observed at 52 kDa.
ab67422 Anti-Folate Binding Protein/FBP antibody was shown to specifically react with Folate Binding Protein/FBP in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264921 (knockout cell lysate ab257270) was used. Wild-type and Folate Binding Protein/FBP knockout samples were subjected to SDS-PAGE. ab67422 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
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Western blot - Anti-Folate Binding Protein/FBP antibody (ab67422)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. -
Immunocytochemistry/ Immunofluorescence - Anti-Folate Binding Protein/FBP antibody (ab67422)
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.
