AB264921

Human FOLR1 (Folate Binding Protein/FBP) knockout HeLa cell line

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FOLR1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 4 bp deletion in exon 1 and Insertion of the selection cassette in exon 1. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.

別名を表示する

Adult folate-binding protein, FBP, FOLR, FOLR1_HUMAN, FR-alpha, Folate Binding Protein, Folate Receptor 1 Adult, Folate Receptor 1 Precursor, Folate receptor, Folate receptor 1, Folate receptor adult, Folate receptor alpha, KB cells FBP, MOV18, Ovarian cancer associated antigen, Ovarian tumor associated antigen, Ovarian tumor-associated antigen MOv18

7 Images

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Western blot - Human FOLR1 (Folate Binding Protein/FBP) knockout HeLa cell line (AB264921)

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° 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.

All lanes:

Western blot - Anti-Folate Binding Protein/FBP antibody (<a href='/products/primary-antibodies/folate-binding-protein-fbp-antibody-ab67422'>ab67422</a>) at 1/500 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

FOLR1 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human FOLR1 (Folate Binding Protein/FBP) knockout HeLa cell line (ab264921)

Lane 3:

JAR cell lysate at 20 µg

Lane 4:

MDA-MB-231 cell lysate at 20 µg

Predicted band size: 30 kDa

Observed band size: 38 kDa

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Lab

Western blot - Human FOLR1 (Folate Binding Protein/FBP) knockout HeLa cell line (AB264921)

Lanes 1-4 : Merged signal (red and green). Green - ab221543 observed at 36, 38 kDa. Red - loading control ab7291 observed at 52 kDa.

ab221543 Anti-Folate Binding Protein/FBP antibody [EPR20277] 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. ab221543 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4° at 1 in 1000 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.

The molecular weight (36,38kDa) observed were knockout validated and consistent with what has been described in the literature (PMID : 23144806).

All lanes:

Western blot - Anti-Folate Binding Protein/FBP antibody [EPR20277] (<a href='/products/primary-antibodies/folate-binding-protein-fbp-antibody-epr20277-ab221543'>ab221543</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human FOLR1 (Folate Binding Protein/FBP) knockout HeLa cell line (ab264921)

Lane 2:

FOLR1 knockout HeLa cell lysate at 20 µg

Lane 3:

JAR cell lysate at 20 µg

Lane 4:

MDA-MB-231 cell lysate at 20 µg

Predicted band size: 30 kDa

Observed band size: 38 kDa,36 kDa

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Sanger Sequencing - Human FOLR1 (Folate Binding Protein/FBP) knockout HeLa cell line (AB264921)

Sanger seq

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Sanger Sequencing - Human FOLR1 (Folate Binding Protein/FBP) knockout HeLa cell line (AB264921)

Allele-2 : Insertion of the selection cassette in exon 1.

Sanger seq

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Sanger Sequencing - Human FOLR1 (Folate Binding Protein/FBP) knockout HeLa cell line (AB264921)

Allele-1 : 4 bp deletion in exon 1.

Sanger seq

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Sanger Sequencing - Human FOLR1 (Folate Binding Protein/FBP) knockout HeLa cell line (AB264921)

Sequencing chromatogram displaying sequence edit in exon 1

Cell Culture - Human FOLR1 (Folate Binding Protein/FBP) knockout HeLa cell line (AB264921)

Cell Culture

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Cell Culture - Human FOLR1 (Folate Binding Protein/FBP) knockout HeLa cell line (AB264921)

Representative images of FOLR1 knockout HeLa cells, low and high confluency examples (top left and right respectively) and wild-type HeLa cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS XL Core microscope.

Immunocytochemistry/ Immunofluorescence - Human FOLR1 (Folate Binding Protein/FBP) knockout HeLa cell line (AB264921)

ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Human FOLR1 (Folate Binding Protein/FBP) knockout HeLa cell line (AB264921)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized FOLR1 KO HeLa (FOLR1 knockout human cervical adenocarcinoma epithelial cell), ab264921 cells labelling Folate Binding Protein/FBP with ab322459 at 1/100 (5.27 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing membranous and cytoplasmic staining in wildtype HeLa cells, showing no staining in FOLR1 knockout HeLa cells (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

Key facts

細胞タイプ

HeLa

生物種

Human

組織

Cervix

製品の状態

Liquid

form

ノックアウト検証方法

Sanger Sequencing,Western blot

ノックアウト変異

Knockout achieved by using CRISPR/Cas9, 4 bp deletion in exon 1 and Insertion of the selection cassette in exon 1

疾病

Adenocarcinoma

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Reactivity data

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製品の詳細

We will provide viable cells that proliferate on revival.

This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.

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製品内容

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出荷温度及び保存条件

遺伝子名

FOLR1

遺伝子編集のタイプ

Knockout

遺伝子編集の方法

CRISPR technology

ノックアウト検証方法

Sanger Sequencing, Western blot

出荷温度

Dry Ice

短期保存温度

-196°C

長期保存温度

-196°C

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取り扱い方法

初回取り扱いガイドライン

Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.

1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO₂. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x10⁴ cells/cm². Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.

継代培養ガイドライン

All seeding densities should be based on cell counts gained by established methods.
A guide seeding density of 2x10⁴ cells/cm² is recommended.
Cells should be passaged when they have achieved 80-90% confluence.

培養培地

DMEM (High Glucose) + 10% FBS

凍結保存培地

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

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補足情報

This supplementary information is collated from multiple sources and compiled automatically.

Folate Binding Protein (FBP) also known as FolR1 protein or FOLR-1 is a significant protein with a mass of about 32-38 kDa. It binds folate with high affinity and facilitates its transport into the cell. FBP can be found in various tissues with notable expression in the placenta kidney and certain epithelial tissues. The protein mediates folate uptake by capturing folates in the extracellular matrix and channeling them through receptor-mediated endocytosis.

Biological function summary

FBP plays a critical role in cellular growth and replication by ensuring sufficient folate supply. It belongs to a family of receptors not forming part of a larger protein complex. FBP's primary function is delivering folate an important B vitamin which acts as a one-carbon donor in DNA synthesis and repair. Optimal folate transport by FBP supports cellular functions particularly in rapidly dividing cells.

Pathways

FBP significantly contributes to the folate-mediated one-carbon metabolic pathway critical for nucleotide biosynthesis. This pathway interconnects with the homocysteine remethylation cycle. FBP associates with other enzymes such as dihydrofolate reductase (DHFR) and methylenetetrahydrofolate reductase (MTHFR) which also play key roles in these pathways. These interactions ensure efficient folate metabolism and maintenance of normal cellular functions.

FBP's relation to cancer and neural tube defects is well documented. Overexpression of FBP has been observed in various cancers including ovarian and breast cancer which indicates its potential role in tumor progression. In neural tube defects altered folate transport due to variations in FBP expression impacts neural development. FBP connects to other markers like HER2 in cancer which may form part of a therapeutic target strategy.

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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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製品プロトコール

Visit the General protocols
Visit the Troubleshooting

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ターゲットの情報

See full target information FOLR1

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Abcam product promise

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Human FOLR1 (Folate Binding Protein/FBP) knockout HeLa cell line