CD229 antibody | HLy9.1.25
Mouse anti Human CD229 antibody, clone HLy9.1.25 used for the detection of SLAMF3 (CD229) using flow cytometry.
Image caption:
SLAMF3 is expressed by hepatocytes. (A) SLAMF3 expression was analysed by flow cytometry analysis in HHPHs and in Huh-7, HepG2 and Hep3B human HCC cell lines. SLAMF3 expression in Jurkat lymphocytes was used as a positive control; SLAMF3 staining (in grey) is overlaid by the negative control (in white) and corresponds to one representative of five independent experiments. (B) Western blot analysis of proteins extracted from Huh-7 cells, HepG2 cells or HHPHs, with a mAb (K12) against SLAMF3′s first extracellular domain (D1) or an anti-actin antibody as a control. One of four independent experiments is presented here. (C) Expression of SLAMF3 transcripts in hepatocytes. After reverse transcription, SLAMF3 cDNA was amplified by PCR using specific primers. GAPDH was amplified as a control gene and pure H2O was used as a PCR control. The Daudi and Jurkat human lymphocyte cell lines were used as positive controls and the monkey kidney COS-7 cell line was used as a negative control. One of three independent experiments is shown here; (D) SLAMF3 mRNA was assayed by Q-PCR in (i) HHPHs, Huh-7 cells and HepG2 cells, (ii) Daudi B lymphocytes and Jurkat T lymphocytes (positive controls) and (iii) the green monkey kidney COS-7 cell line (a negative control). Results are presented as the mean ± SD (n = 6) ***p<0.005, **p<0.01).
From: Marcq I, Nyga R, Cartier F, Amrathlal RS, Ossart C, et al. (2013) Identification of SLAMF3 (CD229) as an Inhibitor of Hepatocellular Carcinoma Cell Proliferation and Tumour Progression. PLoS ONE 8(12): e82918.
Mouse anti Human CD229 antibody, clone HLy9.1.25 used for the detection of SLAMF3 (CD229) using flow cytometry.
Image caption:
Correlation between SLAMF3 expression and HCC cell proliferation. (A) Effects of the introduction of specific siRNAs on SLAMF3 expression in Huh-7 cells. Cells were transiently transfected with a scrambled siRNA (Sc) or one of two pre-designed anti-SLAMF3 siRNAs (referred to as #2 and # 3); SLAMF3 expression was measured by Western blot analysis (anti-SLAMF3, K12). One representative of two independent experiments is shown. (B) siRNA-treated Huh-7 cells were cultured and proliferation was assayed in a Trypan blue exclusion test after 48 h. Proliferation is presented as the mean number of viable cells ± SD (n = 5; statistical significance: ***p<0.005). (C) Following our observation of bimodal SLAMF3 expression, Huh-7 cells were sorted into SLAMF3neg and SLAMF3pos subpopulations, as shown by the dot plot (top panel); SLAMF3 expression in the two enriched populations was measured by flow cytometry (bottom panel); (D) Huh-7 SLAMF3neg and Huh-7 SLAMF3pos cells were cultured separately and cell proliferation was assayed in a Trypan blue exclusion test after 24, 48 and 72 h. Proliferation is presented as mean number of viable cells ± SD (n = 4; statistical significance: **p<0.01, ***p<0.005); SLAMF3 expression was forced in Huh-7 and HepG2 cell lines and proliferation was determined in an MTT assay (E) after 24, 48 and 72 h. The results are presented as the mean ± SD (n = 4; statistical significance: *p<0.05); (F) Investigation of the role of SLAMF3′s first extracellular domain (D1) in the protein’s suppressor effect on HCC proliferation. COS-7 cells (which are naturally negative for SLAMF3) were transfected with a vector coding for wild type SLAMF3 or SLAMF3 lacking D1 (delta-D1 SLAMF3) or a mock pBud vector. Cell proliferation was monitored with MTT assays performed 24 and 72 h after transfection. The results are presented as the mean ± SD (n = 3; statistical significance: *p<0.05, **p<0.01, ***p<0.005; ns: non-significant).
From: Marcq I, Nyga R, Cartier F, Amrathlal RS, Ossart C, et al. (2013) Identification of SLAMF3 (CD229) as an Inhibitor of Hepatocellular Carcinoma Cell Proliferation and Tumour Progression. PLoS ONE 8(12): e82918.
Mouse anti Human CD229 antibody, clone HLy9.1.25 used for the detection of SLAMF3 (CD229) using flow cytometry.
Image caption:
Restoration of SLAMF3 expression in HCC cells inhibits MAPK ERK1/2, JNK and mTOR pathways and induces caspase-dependent apoptosis. (A) SLAMF3 expression was confirmed by Western blot analysis in SLAMF3-over-expressing Huh-7 and mock cells at 48 h post-transfection. ERK1/2, JNK, p38, AKT and mTOR proteins were detected as controls. The activation levels of MAPK ERK1/2 (Thr 202/Tyr 204), JNK (Thr 183/Tyr 185), p38 (Thr 180/Tyr 183), PI3K (p85/p110 Thr 467/Tyr 199), AKT (Ser 473, Thr 308) and mTOR (Ser 2448, Ser 2481) are represented. One of three representative experiments is shown here. (B) The results of an active caspase assay based on cell-permeable fluorochrome inhibitor of caspases (FLICA). Caspase activity was evaluated in HCC cells (Huh-7 and HepG2 lines) at the indicated times after the ectopic introduction of SLAMF3 vector. Caspase activity is shown in the SLAMF3neg subpopulation (in white) and the SLAMF3pos (overlaid in grey) subpopulation from one representative of two independent experiments.
From: Marcq I, Nyga R, Cartier F, Amrathlal RS, Ossart C, et al. (2013) Identification of SLAMF3 (CD229) as an Inhibitor of Hepatocellular Carcinoma Cell Proliferation and Tumour Progression. PLoS ONE 8(12): e82918.
Mouse anti Human CD229 antibody, clone HLy9.1.25 used for the detection of SLAMF3 (CD229) by immunohistochemistry on formalin fixed, paraffin embedded tissue sections.
Image caption:
SLAMF3 expression is repressed in tumour cells from ten resected HCC patients. (A) SLAMF3 mRNA expression was analysed in hepatocytes from resected HCC patients. Specific amplification of SLAMF3 mRNAs using PCR (representative patients #2, #7 and #12) and quantitative PCR in tumour tissues (T) and peritumoral tissues (pT) presented separately (B) and as median (**p<0.05) (C); (D) Immunohistochemical staining of SLAMF3 in T and pT areas from representative patients #3 and #12 (×20).
From: Marcq I, Nyga R, Cartier F, Amrathlal RS, Ossart C, et al. (2013) Identification of SLAMF3 (CD229) as an Inhibitor of Hepatocellular Carcinoma Cell Proliferation and Tumour Progression. PLoS ONE 8(12): e82918.
Mouse anti Human CD229 antibody, clone HLy9.1.25 used for the detection of SLAMF3 (CD229) using flow cytometry.
Image caption:
Expression of SLAM-R family by HHPHs and Huh-7 and HepG2 human HCC cell lines. SLAM-Rs were stained with specific antibodies against SLAMF1 (CD150), SLAMF2 (CD84), SLAMF4 (CD224) and SLAMF6 (NTBA) (grey) or with matched isotype controls (empty). One of four independent experiments is shown.
From: Marcq I, Nyga R, Cartier F, Amrathlal RS, Ossart C, et al. (2013) Identification of SLAMF3 (CD229) as an Inhibitor of Hepatocellular Carcinoma Cell Proliferation and Tumour Progression. PLoS ONE 8(12): e82918.
Mouse anti Human CD229 antibody, clone HLy9.1.25 used for the detection of SLAMF3 (CD229) using flow cytometry.
Image caption:
Expression of SLAMF3 assessed by flow cytometry analysis after transfection with vector coding for SLAMF3 or an empty vector (Mock) in Huh-7 and HepG2 cells. SLAMF3 staining (in grey) is overlaid by negative control (in white). One of four independent experiments is shown. doi:10.1371/journal.pone.0082918.s002
From: Marcq I, Nyga R, Cartier F, Amrathlal RS, Ossart C, et al. (2013) Identification of SLAMF3 (CD229) as an Inhibitor of Hepatocellular Carcinoma Cell Proliferation and Tumour Progression. PLoS ONE 8(12): e82918.
Mouse anti Human CD229 antibody, clone HLy9.1.25 used for the detection of SLAMF3 (CD229) by immunofluorescence.
Image caption:
Effect of SLAMF3 expression on the organization of the actin cytoskeleton. Cells (Huh-7) were stained with phalloidin (rhodamine, red) and anti-SLAMF3 (FITC, green) and SLAMF3 positive (Huh-7-SLAMF3pos) and SLAMF3-negative (Huh-7-SLAMF3pos) cells were examined under the microscope. One representative of two independent experiments is shown.
From: Marcq I, Nyga R, Cartier F, Amrathlal RS, Ossart C, et al. (2013) Identification of SLAMF3 (CD229) as an Inhibitor of Hepatocellular Carcinoma Cell Proliferation and Tumour Progression. PLoS ONE 8(12): e82918.
Mouse anti Human CD229 antibody, clone HLy9.1.25 used for the detection of SLAMF3 (CD229) by immunofluorescence.
Image caption:
SLAMF3 over-expression increases hepatocyte permissiveness to HCV. Huh-7 cells were transfected with empty vector pBud (mock) or human SLAMF3 (to yield Huh-7+ cells) and incubated with HCVcc. (A) SLAMF3 expression was analyzed by flow cytometry (HLy9.1.25 clone) and (B) visualized by confocal microscopy in one out of three experiment. Unshaded histograms show the isotype-matched control (Co) and shaded histograms show SLAMF3 expression). (C, D, E) Infection was assessed at 4 and 72 h p.i. as the number of FFUs and (F and G) by flow cytometry analysis; (C) intracellular E2 (stained red) were measured at 4 h and 72 h p.i. Green staining corresponds to the SLAMF3 expression. One of three representatives independent experiments is shown. (D) The HCV permissiveness of Huh-7 mock and Huh-7+ cells was evaluated in terms of the number of FFUs based on intracellular E2 viral protein staining and (E) fluorescent microscopy analysis (mean of five experiments; error bars: SD; *p<0.05); Infection evaluation by flow cytometry analysis measuring the intracellular viral E2. (F) Infected cells (72 p.i.) were stained for surface expression of SLAMF3 (HLy9.1.25 clone) and permeabilized for intracellular E2 staining. The percentage of positive cells in each cell population is indicated in the corresponding quadrant (one of three representative experiments is shown) and (G) as a mean of three experiments (error bars: SD; *p<0.05).
From: Cartier F, Marcq I, Douam F, Ossart C, Regnier A, et al. (2014) The Expression of the Hepatocyte SLAMF3 (CD229) Receptor Enhances the Hepatitis C Virus Infection.
PLoS ONE 9(6): e99601.
Mouse anti Human CD229 antibody, clone HLy9.1.25 used for the detection of SLAMF3 (CD229) using flow cytometry.
Image caption:
SLAMF3 blockade in human hepatocytes is associated with lower susceptibility to HCV. (A) SLAMF3 was stained in primary human hepatocytes (PHHs) and cells from the Huh-7 human hepatoma cell line with a specific antibody (HLy9.1.25 clone; grey) and an isotype-matched control (empty). One of four independent experiments is shown. Huh-7 cells were transfected with scrambled control (sc) siRNA or three specific siRNAs (#1, #2 and #3) targeting SLAMF3, prior to infection with HCVcc; siRNA efficiency was checked by quantifying SLAMF3 mRNA (B) and the CD81 expression level (C) by flow cytometry analysis at 48 h post-transfection. Results are presented as the mean ±SD (n = 3). Intracellular viral RNA was quantified at 72 h p.i. (D) and the infection was measured at 72 h p.i. by focus-forming units FFUs counting (E) (as inhibition percent; mean of three independent experiments; error bars: SD. **p<0.01);
From: Cartier F, Marcq I, Douam F, Ossart C, Regnier A, et al. (2014) The Expression of the Hepatocyte SLAMF3 (CD229) Receptor Enhances the Hepatitis C Virus Infection. PLoS ONE 9(6): e99601.
Mouse anti Human CD229 antibody, clone HLy9.1.25 recognizes the human cell surface antigen CD229 also known as T-lymphocyte surface antigen Ly-9 or SLAM family member 3. CD229 is a 608 amino acid single pass type I transmembrane glycoprotein of ~120 kDa as evaluated by immunoprecipitation of cells transfected with the full length human CD229 cDNA. However immunoprecipitation of CD229 from Daudi cell lysates with dlone HLy9.1.25 yields bands of 120 kDa corresponding to the full length CD229 and a 100 kDa band attributed to an alternatively spliced isoform lacking the fourth Ig-like domain (de la Fuente et al. 2001).
Human CD299 is expressed on thymocytes, T-cells and B-cells (Del Valle et al. 2003). CD229 has also been described as a tumor associated antigen in chronic lymphocytic leukemia (Bund et al. 2006) and has been implicated in the development of spontaneous autoantibody production to nuclear antigens in mice and is potentially a target for the treatment of autoimmunity (de Salort et al. 2013).
Human CD299 is expressed on thymocytes, T-cells and B-cells (Del Valle et al. 2003). CD229 has also been described as a tumor associated antigen in chronic lymphocytic leukemia (Bund et al. 2006) and has been implicated in the development of spontaneous autoantibody production to nuclear antigens in mice and is potentially a target for the treatment of autoimmunity (de Salort et al. 2013).
Product Details
- Target Species
- Human
- Product Form
- Purified IgG - liquid
- Product Form
- Purified IgG conjugated to R. Phycoerythrin (RPE) - lyophilized
- Reconstitution
- Reconstitute with 1 ml distilled water
- Preparation
- Purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant
- Preparation
- Purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant
- Buffer Solution
- Phosphate buffered saline
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
0.09% Sodium Azide - Preservative Stabilisers
0.09% Sodium Azide 1% Bovine Serum Albumin 5% Sucrose - Carrier Free
- Yes
- Immunogen
- Mouse pre-B cell line 300.19 transfected with CD229 cDNA.
- Approx. Protein Concentrations
- IgG concentration 1.0 mg/ml
- Fusion Partners
- Spleen cells from immunised BALB/c mice were fused with cells of the mouse NS-1 myeloma cell line.
Storage Information
- Storage
- Store at +4oC or at -20oC if preferred.
This product should be stored undiluted.
Storage in frost-free freezers is not recommended. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use. - Storage
- Prior to reconstitution store at +4oC. Following reconstitution store at +4oC.
DO NOT FREEZE.
This product should be stored undiluted. This product is photosensitive and should be protected from light. Should this product contain a precipitate we recommend microcentrifugation before use. - Shelf Life
- 18 months from date of despatch.
- Shelf Life
- 12 months from date of reconstitution.
More Information
- UniProt
- Q9HBG7 Related reagents
- Entrez Gene
- LY9 Related reagents
- GO Terms
- GO:0007155 cell adhesion
- GO:0016021 integral to membrane
- GO:0016064 immunoglobulin mediated immune response
- Regulatory
- For research purposes only
Applications of CD229 antibody
| Application Name | Verified | Min Dilution | Max Dilution |
|---|---|---|---|
| Flow Cytometry | 1/50 | 1/100 | |
| Immunoprecipitation | |||
| Flow Cytometry | Neat |
Where this antibody has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. It is recommended that the user titrates the antibody for use in their own system using appropriate negative/positive controls.
Where this antibody has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. It is recommended that the user titrates the antibody for use in their own system using appropriate negative/positive controls.
- Flow Cytometry
- Use 10ul of the suggested working dilution to label 106 cells in 100ul or 100ul whole blood.
- Flow Cytometry
- Use 10ul of the suggested working dilution to label 106 cells in 100ul or 100ul whole blood.
Copyright © 2018 Bio-Rad Antibodies (formerly AbD Serotec)
Secondary Antibodies Available
Negative Isotype Controls Available
| Description | Product Code | Pack Size | Applications | List Price | Quantity |
|---|---|---|---|---|---|
| Mouse IgG1 Negative Control | MCA928 | 100 Tests | F | ||
| Mouse IgG1 Negative Control:RPE | MCA928PE | 100 Tests | F |
Useful Reagents Available
| Description | Product Code | Pack Size | Applications | List Price | Quantity |
|---|---|---|---|---|---|
| Human Seroblock | BUF070A | 50 Test | F | ||
| Human Seroblock | BUF070B | 200 Test | F |
Product Specific References
References for CD229 antibody
-
de la Fuente, M.A. et al. (2001) Molecular characterization and expression of a novel human leukocyte cell-surface marker homologous to mouse Ly-9.
Blood. 97 (11): 3513-20. -
Sayós J et al. (2001) Cell surface receptors Ly-9 and CD84 recruit the X-linked lymphoproliferative disease gene product SAP.
Blood. 97 (12): 3867-74. -
Cartier, F. et al. (2014) The expression of the hepatocyte SLAMF3 (CD229) receptor enhances the hepatitis C virus infection.
PLoS One. 9: e99601. -
Marcq, I. et al. (2013) Identification of SLAMF3 (CD229) as an Inhibitor of Hepatocellular Carcinoma Cell Proliferation and Tumour Progression
PLoS One. 8: e82918. -
Margraf, S. et al. (2015) A polymorphism in a phosphotyrosine signalling motif of CD229 (Ly9, SLAMF3) alters SH2 domain binding and T-cell activation.
Immunology. 146 (3): 392-400.
