CD200R1L antibody (AA 151-250)

Catalog No. ABIN1715098

Product Details anti-CD200R1L Antibody

Target: CD200R1L (CD200 Receptor 1-Like (CD200R1L))

Binding Specificity: AA 151-250

Reactivity: Human

Host: Rabbit

Clonality: Polyclonal

Conjugate: This CD200R1L antibody is un-conjugated

Application: Western Blotting (WB), ELISA, Flow Cytometry (FACS), Immunofluorescence (Cultured Cells) (IF (cc)), Immunofluorescence (Paraffin-embedded Sections) (IF (p)), Immunohistochemistry (Frozen Sections) (IHC (fro)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunocytochemistry (ICC)

Cross-Reactivity: Human

Purification: Purified by Protein A.

Immunogen: KLH conjugated synthetic peptide derived from human CD200R2

Isotype: IgG

Application Details

Application Notes: 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
ICC 1:100-500

Restrictions: For Research Use only

Independent Validation (IHC)

Validation #101354 (Immunohistochemistry)

by: Department of Pathology and Molecular Medicine, McMaster University

No.: #101354

Date: 06/01/2017

Antigen: CD200R1L

Lot Number: 9A13M60

Method validated: Immunohistochemistry

Positive Control: Placental villus from 7w gestation karyotype-normal pregnancy

Negative Control: KLH antibody ABIN401183

Notes: Passed. ABIN1715098 specifically labels the CD200R1L in human placental villus trophoblast tissue in IHC.

Validation Images

Representative villus from a 7 week gestation normal pregnancy with “normal karyotype” as determined by PCR testing to exclude trisomy, and digitally photographed at 400x. Staining with CD200R1L antibody ABIN1715098 (A) or KLH antibody ABIN401183 (B). VC: villus core, CT: cytotrophoblast, ST: syncytiotrophoblast, MB: maternal blood space bathing placental villi. Note more intense staining of cytotrophoblasts compared to syncytiotrophoblasts. Bars represent 60µm.

Full Methods

Primary Antibody: ABIN1715098

Secondary Antibody: Dako EnVision-HRP anti-rabbit polymer (Dako, K4003, lot 10095566)

Full Protocol:

• Human placental tissue is fixed in 10% buffered formalin for 24h at RT and embedded in paraffin.
• Cut paraffin blocks with a Leica 12M2255 Microtome into 4µm sections.
• Affix sections to positively charged slides and air dry.
• Deparaffinize and rehydrate sections through graded xylene and graded alcohol series:
• • xylene 3x 5 min.
  • 100% ethanol 3x 1min.
  • 95% ethanol for 1min.
  • 70% ethanol for 1min.
  • dH₂O for 2min.
• Heat the sections in the decloaking Digital Decloaking chamber (Biacore, DC-2002) using factory settings (125°C for 30sec and cool) in 1mM EDTA buffer pH8.0 (0.37g EDTA (disodium salt) in 1l dH₂O, adjust to pH8.0 using 1 M NaOH, add 1 small crystal thymol).
• Rinse sections 3 times with 0.05 M Tris-buffered saline pH 7.6 with 0.1% Tween 20 (TBST) 3x for 5min.
• Block sections in 5% normal goat serum (Jackson Immuno Research, 005-000-121, lot 116831) in TBST fo 30min at RT.
• Blot excess serum from sections.
• Incubate sections with
• • primary rabbit anti-CD200 Receptor 1-Like (CD200R1L) (AA 150-200) antibody(antibodies-online, ABIN1715098, lot 9A13M60) diluted 1:250 in antibody diluent (Thermo Fisher TA-125-ADQ Lot ADQ 151111) ON at 4°C.
  • primary rabbit anti-KLH antibody also antigen-affinity purified (antibodies-online, ABIN401183, lot 30477) diluted 1:250 in Antibody Diluent (as above) ON at 4°C.
• Rinse sections 3x 5min with TBST.
• Stain sections with Dako EnVision-HRP anti-rabbit polymer (Dako, K4003, lot 10095566) for 30min at RT.
• Rinse sections 3x 5min with TBST.
• Stain 30min with AEC (5 ml AEC stock (80mg AEC in 20ml Dimethylformamide), 95 ml 0.05M acetate buffer pH 5.0, filter and add 2 drops of 30% hydrogen peroxide to activate).
• Rinse with pH5.0 acetate buffer.
• Counterstain 1min with 50% Mayer’s hematoxylin.
• Wash in tap water and blue 1min in TBST.
• Mount sections in 15% glycerine/gelatin in TBST pH 8.0 and apply coverslip. Glycerine solution is made with 50 g gelatin (100 bloom) in 150ml TBS pH 8.0, incubated in a 60°C oven until the gelatin is melted, the 175 ml glycerine and a small pellet of thymol is added.
• Allow sections to dry.
• Scan slides using Imagescope and photograph at 400x. Quantification using Image J 1.4.

Experimental Notes: The result shown is validated by the data of Wang et al. (2014) Acta Obstetrica et Gynecologica Scandinavica where a non-antigen-affinity-purified goat polyclonal antibody to CD200R1 and no negative control antibody was used. Wang et al. confirmed expression of CD200R1 using real time PCR of villus mRNA.

Handling

Format: Liquid

Concentration: 1 μg/μL

Buffer: 0.01M TBS( pH 7.4) with 1 % BSA, 0.02 % Proclin300 and 50 % Glycerol.

Preservative: ProClin

Precaution of Use: This product contains ProClin: a POISONOUS AND HAZARDOUS SUBSTANCE, which should be handled by trained staff only.

Storage: 4 °C,-20 °C

Storage Comment: Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.

Expiry Date: 12 months

References for anti-CD200R1L antibody (ABIN1715098)

Hamilton, Turpin, Ayoub, Reihani, Arredondo, Ask, Clark, Foster: "Circulating CD200 is increased in the secretory phase of women with endometriosis as is endometrial mRNA, and endometrial stromal cell CD200R1 is increased in spite of reduced mRNA." in: American journal of reproductive immunology (New York, N.Y. : 1989), Vol. 89, Issue 1, pp. e13655, (2022) (PubMed).

Clark, Dmetrichuk, McCready, Dhesy-Thind, Arredondo: "Changes in expression of the CD200 tolerance-signaling molecule and its receptor (CD200R) by villus trophoblasts during first trimester missed abortion and in chronic histiocytic intervillositis." in: American journal of reproductive immunology (New York, N.Y. : 1989), Vol. 78, Issue 1, (2017) (PubMed).

Clark, Dhesy-Thind, Arredondo, Ellis, Ramsay: "The Receptor for the CD200 Tolerance-Signaling Molecule Associated with Successful Pregnancy is Expressed by Early-Stage Breast Cancer Cells in 80% of Patients and by Term Placental Trophoblasts." in: American journal of reproductive immunology (New York, N.Y. : 1989), Vol. 74, Issue 5, pp. 387-91, (2015) (PubMed).

Clark, Arredondo, Dhesy-Thind: "The CD200 tolerance-signaling molecule and its receptor, CD200R1, are expressed in human placental villus trophoblast and in peri-implant decidua by 5 weeks' gestation." in: Journal of reproductive immunology, Vol. 112, pp. 20-3, (2015) (PubMed).

Target Details for CD200R1L

Target: CD200R1L (CD200 Receptor 1-Like (CD200R1L))

Alternative Name: CD200R2 (CD200R1L Products)

Synonyms: CD200R2 antibody, CD200RLa antibody, AY230198 antibody, Cd200r1l antibody, CD200 receptor 1 like antibody, Cd200 receptor 2 antibody, CD200R1L antibody, Cd200r2 antibody

Background:

Synonyms: CD200R2, CD200RLa, Cell surface glycoprotein CD200 receptor 2, CD200 cell surface glycoprotein receptor-like 2, CD200 receptor-like 2, HuCD200R2, CD200 cell surface glycoprotein receptor-like a, Cell surface glycoprotein CD200 receptor 1-like, Cell surface glycoprotein OX2 receptor 2, CD200R1L

Background: May be a receptor for the CD200/OX2 cell surface glycoprotein.

Gene ID: 344807

UniProt: Q6Q8B3