Human FAM20B Antibody (1018512)

R&D Systems | Catalog # MAB8427

R&D Systems

Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Western Blot

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 1018512
View all FAM20B Primary Antibodies »

Product Specifications

Immunogen

Chinese hamster ovary cell line CHO-derived recombinant human FAM20B
Asn27-Leu409
Accession # O75063

Specificity

Detects human FAM20B in direct ELISAs.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Scientific Data Images for Human FAM20B Antibody (1018512)

Detection of Human FAM20B antibody by Western Blot.

Detection of Human FAM20B by Western Blot.

Western blot shows lysates of ZR-75 human breast cancer cell line. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human FAM20B Monoclonal Antibody (Catalog # MAB8427) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for FAM20B at approximately 46 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of FAM20B by Western Blot

Detection of FAM20B by Western Blot

FAM20C-mediated control of CS biosynthesis is exerted by its physical interaction with C4ST-1.a Schematic diagram of sulfation pathways for the chondroitin backbone. Characteristic CS disaccharide units, including A, C, D, and E, are sequentially formed under the control of CS-specific sulfotransferases such as C4ST-1, C4ST-2, and C6ST-1. b Gel filtration elution profiles of CS chains from mouse fibroblastic L or sog9 cells overexpressing wild-type FAM20B or FAM20C (Data are derived from the single experiment). Numbered arrowheads (200 k, 65 k, 37 k, and 18 k) indicate the elution position of 200-, 65-, 37-, and 18-kDa saccharides derived from size-defined commercial dextran, respectively. c Pulldown (PD) and immunoblotting (IB) analysis of culture medium from COS-1 cells co-expressing soluble forms of His-tagged C4ST-1 and FLAG-tagged FAM20 proteins. Data are obtained from three independent experiments and representative images are shown. d Sulfotransferase activities of C4ST-1 alone (Mock) and that from cells co-transfected with the individual FAM20 proteins (n = 3 independent experiments, Dunnett’s multiple comparison test, two-sided). e The 4S/6S ratio of CS chains from HeLa cell lines overexpressing wild-type or mutant FAM20C proteins (n = 4 independent experiments, Dunnett’s multiple comparison test, two-sided). Data in d and e are represented as the mean ± s.d. Source data are provided as a Source Data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36572689), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of FAM20B by Western Blot

Detection of FAM20B by Western Blot

FAM20C mutants enhance biomineralization in human osteosarcoma cells. A, b Expression of FAM20C, FAM20B, and C4ST-1 in stable Saos-2 cell lines overexpressing individual FAM20 proteins at the a transcriptional and b translational levels. GAPDH was used as an internal standard in (a), and as a loading control in (b). Data are obtained from three independent experiments and representative images are shown. c The 4S/6S ratio of CS chains from the stable Saos-2 cell lines in (a) and (b) (n = 3 independent experiments, Dunnett’s multiple comparison test, two-sided). d, e Biomineralization level of the 21-day cultures of the stable Saos-2 cell lines in (a) and (b) was assessed by Alizarin red staining (d), followed by colorimetric quantification of the dye extracts derived from the stained cultures (e, n = 4 independent experiments, Dunnett’s multiple comparison test, two-sided). Data in c and e are represented as the mean ± s.d. Source data are provided as a Source Data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36572689), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of FAM20B by Western Blot

Detection of FAM20B by Western Blot

FAM20C mutants enhance biomineralization in human osteosarcoma cells. A, b Expression of FAM20C, FAM20B, and C4ST-1 in stable Saos-2 cell lines overexpressing individual FAM20 proteins at the a transcriptional and b translational levels. GAPDH was used as an internal standard in (a), and as a loading control in (b). Data are obtained from three independent experiments and representative images are shown. c The 4S/6S ratio of CS chains from the stable Saos-2 cell lines in (a) and (b) (n = 3 independent experiments, Dunnett’s multiple comparison test, two-sided). d, e Biomineralization level of the 21-day cultures of the stable Saos-2 cell lines in (a) and (b) was assessed by Alizarin red staining (d), followed by colorimetric quantification of the dye extracts derived from the stained cultures (e, n = 4 independent experiments, Dunnett’s multiple comparison test, two-sided). Data in c and e are represented as the mean ± s.d. Source data are provided as a Source Data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36572689), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of FAM20B by Western Blot

Detection of FAM20B by Western Blot

FAM20C mutants enhance biomineralization in human osteosarcoma cells. A, b Expression of FAM20C, FAM20B, and C4ST-1 in stable Saos-2 cell lines overexpressing individual FAM20 proteins at the a transcriptional and b translational levels. GAPDH was used as an internal standard in (a), and as a loading control in (b). Data are obtained from three independent experiments and representative images are shown. c The 4S/6S ratio of CS chains from the stable Saos-2 cell lines in (a) and (b) (n = 3 independent experiments, Dunnett’s multiple comparison test, two-sided). d, e Biomineralization level of the 21-day cultures of the stable Saos-2 cell lines in (a) and (b) was assessed by Alizarin red staining (d), followed by colorimetric quantification of the dye extracts derived from the stained cultures (e, n = 4 independent experiments, Dunnett’s multiple comparison test, two-sided). Data in c and e are represented as the mean ± s.d. Source data are provided as a Source Data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36572689), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of FAM20B by Western Blot

Detection of FAM20B by Western Blot

FAM20C mutants enhance biomineralization in human osteosarcoma cells. A, b Expression of FAM20C, FAM20B, and C4ST-1 in stable Saos-2 cell lines overexpressing individual FAM20 proteins at the a transcriptional and b translational levels. GAPDH was used as an internal standard in (a), and as a loading control in (b). Data are obtained from three independent experiments and representative images are shown. c The 4S/6S ratio of CS chains from the stable Saos-2 cell lines in (a) and (b) (n = 3 independent experiments, Dunnett’s multiple comparison test, two-sided). d, e Biomineralization level of the 21-day cultures of the stable Saos-2 cell lines in (a) and (b) was assessed by Alizarin red staining (d), followed by colorimetric quantification of the dye extracts derived from the stained cultures (e, n = 4 independent experiments, Dunnett’s multiple comparison test, two-sided). Data in c and e are represented as the mean ± s.d. Source data are provided as a Source Data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36572689), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of FAM20B by Western Blot

Detection of FAM20B by Western Blot

FAM20C-mediated control of CS biosynthesis is exerted by its physical interaction with C4ST-1.a Schematic diagram of sulfation pathways for the chondroitin backbone. Characteristic CS disaccharide units, including A, C, D, and E, are sequentially formed under the control of CS-specific sulfotransferases such as C4ST-1, C4ST-2, and C6ST-1. b Gel filtration elution profiles of CS chains from mouse fibroblastic L or sog9 cells overexpressing wild-type FAM20B or FAM20C (Data are derived from the single experiment). Numbered arrowheads (200 k, 65 k, 37 k, and 18 k) indicate the elution position of 200-, 65-, 37-, and 18-kDa saccharides derived from size-defined commercial dextran, respectively. c Pulldown (PD) and immunoblotting (IB) analysis of culture medium from COS-1 cells co-expressing soluble forms of His-tagged C4ST-1 and FLAG-tagged FAM20 proteins. Data are obtained from three independent experiments and representative images are shown. d Sulfotransferase activities of C4ST-1 alone (Mock) and that from cells co-transfected with the individual FAM20 proteins (n = 3 independent experiments, Dunnett’s multiple comparison test, two-sided). e The 4S/6S ratio of CS chains from HeLa cell lines overexpressing wild-type or mutant FAM20C proteins (n = 4 independent experiments, Dunnett’s multiple comparison test, two-sided). Data in d and e are represented as the mean ± s.d. Source data are provided as a Source Data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36572689), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human FAM20B Antibody (1018512)

Application
Recommended Usage

Western Blot

2 µg/mL
Sample: ZR‑75 human breast cancer cell line

Reviewed Applications

Read 1 review rated 5 using MAB8427 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: FAM20B

FAM20B is a member of the FAM20 protein family that has three members in mammals (FAM20A, FAM20B, FAM20C) (1). Human FAM20B shares 97% amino acid sequence identity with mouse and rat FAM20B. FAM20B localizes to the Golgi apparatus where it phosphorylates the xylose residue in the glycosaminoglycan (GAG)-protein linkage region of proteoglycans, which leads to enhanced GAG biosynthesis (2). Accordingly, chondroitin sulfate and heparan sulfate production is increased when FAM20B is over-expressed and reduced following FAM20B knockdown (2). FAM20B knockout mice display embryonic lethality at day E13.5, suggesting that FAM20B has an important role during development (3). Furthermore, in zebrafish, FAM20B mutations result in reduced cartilage matrix production and skeletal defects (4). The enzymatic activity of recombinant human FAM20B is measured using a phosphatase-coupled method (5).

References

  1. Nalbant, D. et al. (2005) BMC Genomics 6:11.
  2. Koike, T. et al. (2009) Biochem. J. 421:157.
  3. Vogel, P. et al. (2012) Vet. Pathol. 49:998.
  4. Eames, B.F. et al. (2011) PLoS Genet. 7:e1002246.
  5. Wu, Z.L. (2011) PLoS One 6:e23172.

Long Name

Family with Sequence Similarity 20, Member B

Alternate Names

Glycosaminoglycan Xylosylkinase, gxk1, XYLK, Xylose Kinase

Entrez Gene IDs

9917 (Human); 215015 (Mouse); 304885 (Rat)

Gene Symbol

FAM20B

UniProt

O75063

Additional FAM20B Products

Product Documents for Human FAM20B Antibody (1018512)

Certificate of Analysis

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Product Specific Notices for Human FAM20B Antibody (1018512)

For research use only

Citations for Human FAM20B Antibody (1018512)

Customer Reviews for Human FAM20B Antibody (1018512) (1)

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  • Human FAM20B Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: ZR-75 human breast cancer cell line
    Species: Human
    Verified Customer | Posted 01/29/2022
    Human FAM20B Antibody (1018512) MAB8427

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