BTLA/CD272 Antibody

Western blot shows lysates of mouse spleen tissue and mouse lymph node tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-Mouse BTLA Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3007) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for BTLA at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Activation of PPAR beta /δ results in upregulation of FABP5. (a) PC3M cells were treated with GW0742 (1 μM, 4 h). Expression of FABP5, PDK1, ADRP, and VEGF were assessed by Q-PCR. The data were normalized to the untreated control. (b, c) Denoted cells were treated with 1 μM (a) or 2 μM (b) GW0742 for 16 h. Left panels, immunoblots of FABP5 in untreated versus GW0742-treated cells. Right panel: Densitometry analyses depicting changes in FABP5 expression upon treatment with GW0742 in three independent experiments (mean ± S.D.). (d, e) PC3M cells were transfected with vectors harboring SiScramble or PPAR beta /δ siRNA. Three days later, expression of FABP5, PPAR beta /δ, and VEGF mRNA in were measured by Q-PCR (d) and FABP5 expression was assessed by immunoblot (e). (f) PC3M cells were stably transfected with an empty vector (PGIPZ), or a vector harbouring shFABP5. Expression levels of FABP5, PPAR beta /δ, and VEGF mRNA were measured by Q-PCR. *P < .05 versus nontreated controls. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/20847935), licensed under a CC-BY license. Not internally tested by R&D Systems.

Prostate cancer progression is accompanied by up regulation of FABP5/PPAR beta /δ expression and signalling. (a) Bottom: Expression levels of FABP5 mRNA in denoted cell lines were measured by Q-PCR. Top: Level of FABP5 protein in denoted cell lines assessed by immunoblots. (b) Bottom: Expression levels of PDK-1 and PPAR beta /δ mRNA in denoted cell lines were measured by Q-PCR. Top: Immunoblots of PDK1 in denoted cell lines. Data are mean ± S.D. (n = 3). *P < .02 versus 22Rv.1. (Paired T test). Immunoblots were repeated three times with similar results. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/20847935), licensed under a CC-BY license. Not internally tested by R&D Systems.

Activation of PPAR beta /δ results in upregulation of FABP5. (a) PC3M cells were treated with GW0742 (1 μM, 4 h). Expression of FABP5, PDK1, ADRP, and VEGF were assessed by Q-PCR. The data were normalized to the untreated control. (b, c) Denoted cells were treated with 1 μM (a) or 2 μM (b) GW0742 for 16 h. Left panels, immunoblots of FABP5 in untreated versus GW0742-treated cells. Right panel: Densitometry analyses depicting changes in FABP5 expression upon treatment with GW0742 in three independent experiments (mean ± S.D.). (d, e) PC3M cells were transfected with vectors harboring SiScramble or PPAR beta /δ siRNA. Three days later, expression of FABP5, PPAR beta /δ, and VEGF mRNA in were measured by Q-PCR (d) and FABP5 expression was assessed by immunoblot (e). (f) PC3M cells were stably transfected with an empty vector (PGIPZ), or a vector harbouring shFABP5. Expression levels of FABP5, PPAR beta /δ, and VEGF mRNA were measured by Q-PCR. *P < .05 versus nontreated controls. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/20847935), licensed under a CC-BY license. Not internally tested by R&D Systems.

The FABP5/PPAR beta /δ pathway enhances prostate cancer cell proliferation and transformation. (a) PC3M cells that stably express shFABP5 were cultured in a 24 well plate (2500 cells/well) and treated with vehicle or GW 0742 (1 μM). Cells were counted at the denoted days. Data are mean ± SEM (n = 3). Inset: immunoblotting demonstrating low FABP5 level in shFABP5-expressing. (b) PC3M cells were transfected with a vector harboring PPAR beta /δsiRNA. Four days later, were cultured in a 24 well plate (2500 cells/well) and treated with vehicle or GW 0742 (1 μM). Cells were counted at the denoted days. Inset: Q-PCR analyses demonstrating low PPAR beta /δ in cells transfected with SiRNA towards the receptor. (c) Colony formation assays were conducted in the absence or presence of GW0742 (1 μM). Cells were cultured in 2% agarose for 29 days (see Materials and methods for details). Media was replenished every 4 days. Colonies were visualized by staining with 0.005% crystal violet and counted under a light microscope. Data are mean ± SEM (n = 3). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/20847935), licensed under a CC-BY license. Not internally tested by R&D Systems.

Activation of PPAR beta /δ results in upregulation of FABP5. (a) PC3M cells were treated with GW0742 (1 μM, 4 h). Expression of FABP5, PDK1, ADRP, and VEGF were assessed by Q-PCR. The data were normalized to the untreated control. (b, c) Denoted cells were treated with 1 μM (a) or 2 μM (b) GW0742 for 16 h. Left panels, immunoblots of FABP5 in untreated versus GW0742-treated cells. Right panel: Densitometry analyses depicting changes in FABP5 expression upon treatment with GW0742 in three independent experiments (mean ± S.D.). (d, e) PC3M cells were transfected with vectors harboring SiScramble or PPAR beta /δ siRNA. Three days later, expression of FABP5, PPAR beta /δ, and VEGF mRNA in were measured by Q-PCR (d) and FABP5 expression was assessed by immunoblot (e). (f) PC3M cells were stably transfected with an empty vector (PGIPZ), or a vector harbouring shFABP5. Expression levels of FABP5, PPAR beta /δ, and VEGF mRNA were measured by Q-PCR. *P < .05 versus nontreated controls. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/20847935), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Mu
• Applications: WB
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

BTLA/CD272 Antibody Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant mouse BTLA
  Glu30-Gly176 (Pro41Glu, Thr45Asn, Thr47Lys, Gln52His, Arg55Trp, Gln63Glu, Cys85Trp)
  Accession # Q32MV9
• Specificity: Detects mouse BTLA in direct ELISAs and Western blots. In direct ELISAs and Western blots, less than 1% cross-reactivity with recombinant human BTLA is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: BTLA
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for BTLA/CD272 Antibody

• B and T lymphocyte associated
• B and T lymphocyte attenuator
• B- and T-lymphocyte attenuator
• B- and T-lymphocyte-associated protein
• BTLA
• BTLA1
• CD272 antigen
• CD272
• FLJ16065
• MGC129743

Background

B- and T-lymphocyte attenuator (BTLA; CD272) is a 70 kDa, Ig-superfamily, type I transmembrane glycoprotein that is structurally similar to the CD28 family of T cell co-stimulatory or coinhibitory molecules (1‑3). Unlike CD28 family members, however, the BTLA extracellular Ig domain is an I-type rather than a V-type domain, and BTLA does not form homodimers (4). BTLA also differs from CD28 family members through the interaction of its Ig domain with the TNF superfamily member HVEM (herpesvirus entry mediator; TNFSF14) rather than with B7 family ligands (5). BTLA is a coinhibitory molecule expressed on T cells, B cells and, depending on the mouse strain, macrophages, dendritic and NK cells (6). Expression is low in naïve T cells and increased during antigen-specific induction of anergy. In B cells, BTLA is highest when cells are mature and naïve (6). BTLA apparently limits T cell numbers, since deletion of BTLA results in overproduction of T cells, especially CD8⁺ memory T cells that are hyper-responsive to TCR crosslinking (7). The 305 amino acid (aa) BTLA contains a 29 aa signal sequence, a 153 aa extracellular domain (ECD), a 21 aa transmembrane sequence, and a 102 aa cytoplasmic domain. There are two ITIM motifs and three Tyr phosphorylation sites in the cytoplasmic tail that mediate inhibitory signaling (8, 9). The binding of the BTLA to HVEM does not preclude additional binding of a mammalian stimulatory HVEM ligand, either LIGHT or lymphotoxin-alpha to the complex (4). At least three alleles varying by up to ten extracellular amino acids occur in different mouse strains (6). The ECD of C57BL/6 BTLA shows 51%, 77% and 40% aa identity to that of human, rat and canine BTLA, respectively. A splice variant lacking the Ig domain, termed BTLAs, has been reported (3).

1. Murphy, K. M. et al. (2006) Nat. Rev. Immunol. 6:671.
2. Croft, M. (2005) Trends Immunol. 26:292.
3. Watanabe, N. et al. (2003) Nat. Immunol. 4:670.
4. Compaan, D. M. et al. (2005) J. Biol. Chem. 280:39553.
5. Sedy, J. R. et al. (2005) Nat. Immunol. 6:90.
6. Hurchla, M. A. et al. (2005) J. Immunol. 174:3377.
7. Krieg, C. et al. (2007) Nat. Immunol. 8:162.
8. Gavrieli, M. et al. (2003) Biochem. Biophys. Res. Commun. 312:1236.
9. Chemnitz, J. M. et al. (2006) J. Immunol. 176:6603.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for BTLA/CD272 Antibody (AF3007)(2)

Publications using AF3007

• J Celis-Guti, P Blattmann, Y Zhai, N Jarmuzynsk, K Ruminski, C Grégoire, Y Ounoughene, F Fiore, R Aebersold, R Roncagalli, M Gstaiger, B Malissen Quantitative Interactomics in Primary T Cells Provides a Rationale for Concomitant PD-1 and BTLA Coinhibitor Blockade in Cancer Immunotherapy Cell Rep, 2019-06-11;27(11):3315-3330.e7. 2019-06-11 [PMID: 31189114] (Western Blot, Mouse)
• Elwin Morgan, Padmamalini Kannan-Thulasiraman, Noa Noy Involvement of Fatty Acid Binding Protein 5 and PPAR beta /δ in Prostate Cancer Cell Growth PPAR Research 2010-08-19 [PMID: 20847935]

Bioinformatics

• Gene Symbol: BTLA
• Uniprot:
  • Q32MV9()