bs-2930R
[Primary Antibody]
Frizzled 5/8 Polyclonal Antibody
DATASHEET
Host: Rabbit
Target Protein: Frizzled 5/8
Immunogen Range: 51-150/585
Clonality: Polyclonal
Isotype: IgG
Entrez Gene: 7855, 8325
Source: KLH conjugated synthetic peptide derived from human Frizzled 5
Purification: Purified by Protein A.
Storage Buffer: 0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
Storage: Shipped at 4C. Store at -20C for one year. Avoid repeated freeze/thaw cycles.
Background:
Members of the 'frizzled' gene family encode 7-transmembrane domain proteins that are receptors for Wnt signaling proteins. The Frizzled 5 protein is believed to be the receptor for the Wnt5A ligand. Frizzled 5 has been reported to be expressed in fetal kidney, fetal and adult liver, fetal lung, and adult pancreas. ESTs have been isolated from bone, liver/spleen, placenta, and prostate libraries. Frizzled 5 was cloned from a retina cDNA library. Receptor for Wnt proteins. Component of the Wnt-Fzd-LRP5-LRP6 complex that triggers beta-catenin signaling through inducing aggregation of receptor-ligand complexes into ribosome-sized signalosomes. The beta-catenin canonical signaling pathway leads to the activation of disheveled proteins, inhibition of GSK-3 kinase, nuclear accumulation of beta-catenin and activation of Wnt target genes. A second signaling pathway involving PKC and calcium fluxes has been seen for some family members, but it is not yet clear if it represents a distinct pathway or if it can be integrated in the canonical pathway, as PKC seems to be required for Wnt-mediated inactivation of GSK-3 kinase. Both pathways seem to involve interactions with G-proteins. May be involved in transduction and intercellular transmission of polarity information during tissue morphogenesis and/or in differentiated tissues. Coreceptor along with RYK of Wnt proteins, such as WNT1.
Size: 100ul
Concentration: 1ug/ul
Applications:
WB(1:300-5000)
ELISA(1:500-1000)
FCM(1:20-100)
IHC-P(1:200-400)
IHC-F(1:100-500)
IF(IHC-P)(1:50-200)
IF(IHC-F)(1:50-200)
IF(ICC)(1:50-200)
Predicted Molecular Weight: 62
Cross Reactive Species:
Human
Mouse
Rat
Predicted Cross Reactive Species:
Dog
Cow
Pig
Rabbit
For research use only. Not intended for diagnostic or therapeutic use.
VALIDATION IMAGES
L1 rat lung lysates, L2 rat brain lysates probed (bs-2930R) Anti-Frizzled-5 Polyclonal, Unconjugated at 1:200 in 4˚C. Followed by conjugation to secondary antibody (bs-0295G-HRP) at 1:3000 90min in RT. Predicted band and observed band size: 62kD.
Formalin-fixed and paraffin embedded mouse embryo labeled with Anti-Frizzled 5/8 Polyclonal Antibody, Unconjugated (bs-2930R) at 1:200 followed by conjugation to the secondary antibody and DAB staining.
Formalin-fixed and paraffin embedded mouse pancreas labeled with Anti-Frizzled 5/8 Polyclonal Antibody, Unconjugated (bs-2930R) at 1:200 followed by conjugation to the secondary antibody and DAB staining.
K562 cell lysates probed with Rabbit Anti-Frizzled 5/8 Polyclonal Antibody, Unconjugated (bs-2930R) at 1:300 overnight at 4˚C. Followed by conjugation to secondary antibody (bs-0295G-HRP) at 1:500 for 90 min at RT.
Lane 1: Cerebrum lysates probed with Frizzled 5/8 Polyclonal Antibody, Unconjugated (bs-2930R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
K562 cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 0.1% PBST for 20 min at room temperature, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with Frizzled 5 Antibody(bs-2930R)at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).
Lane 1: Mouse Liver Lysates; Lane 2: Mouse Brain Lysates; Lane 3: Human HepG2 cell Lysates; Lane 4: Human K562 cell Lysates; Lane 5: Human A549 cell Lysates. Probed with Frizzled 5 polyclonal Antibody, unconjugated (bs-2930R) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
Lane 1: Mouse Cerebrum tissue lysates; Lane 2: Rat Cerebrum tissue lysates; Lane 3: Human HepG2 cell lysates; Lane 4: Human 293T cell lysates; Lane 5: Human A549 cell lysates; Lane 6: Human K562 cell lysates; Lane 7: Human HeLa cell lysates; Lane 8: Human Jurkat cell lysates probed with Frizzled 5 Polyclonal Antibody, Unconjugated (bs-2930R) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at room temperature for 60 min.
Paraformaldehyde-fixed, paraffin embedded (mouse liver); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Incubation with (Frizzled 5) Polyclonal Antibody, Unconjugated (bs-2930R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat liver); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Incubation with (Frizzled 5) Polyclonal Antibody, Unconjugated (bs-2930R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human liver carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Incubation with (Frizzled 5) Polyclonal Antibody, Unconjugated (bs-2930R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat colon); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Incubation with (Frizzled 5) Polyclonal Antibody, Unconjugated (bs-2930R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse colon); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Incubation with (Frizzled 5) Polyclonal Antibody, Unconjugated (bs-2930R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Incubation with (Frizzled 5) Polyclonal Antibody, Unconjugated (bs-2930R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Incubation with (Frizzled 5) Polyclonal Antibody, Unconjugated (bs-2930R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human pancreas); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Incubation with (Frizzled 5) Polyclonal Antibody, Unconjugated (bs-2930R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse pancreas); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Incubation with (Frizzled 5) Polyclonal Antibody, Unconjugated (bs-2930R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat pancreas); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Incubation with (Frizzled 5) Polyclonal Antibody, Unconjugated (bs-2930R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human intestine); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Incubation with (Frizzled 5) Polyclonal Antibody, Unconjugated (bs-2930R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.