CD200 antibody (AA 41-140)

Catalog No. ABIN761396

The Rabbit Polyclonal anti-CD200 antibody has been validated for WB, ELISA, IHC (fro), IF (cc), IF (p) and IHC (p) and has been independently validated. It is suitable to detect CD200 in samples from Human. There is 1 publication available.

Quick Overview for CD200 antibody (AA 41-140) (ABIN761396)

Target: CD200

Reactivity: Human

Host: Rabbit

Clonality: Polyclonal

Conjugate: This CD200 antibody is un-conjugated

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

Product Details anti-CD200 Antibody

Binding Specificity: AA 41-140

Cross-Reactivity: Human

Predicted Reactivity: Mouse,Rat

Purification: Purified by Protein A.

Immunogen: KLH conjugated synthetic peptide derived from human CD200

Isotype: IgG

Application Details

Application Notes: WB 1:300-5000
ELISA 1:500-1000
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

Restrictions: For Research Use only

Independent Validation (IHC)

Validation #101517 (Immunohistochemistry)

by: Department of Pathology and Molecular Medicine, McMaster University

No.: #101517

Date: 07/11/2017

Antigen: CD200

Lot Number: AG06145442

Method validated: Immunohistochemistry

Positive Control: Human 7 week normal karyotype elective termination placenta

Negative Control: Placental villus from 7w gestation karyotype-normal pregnancy, stained with KLH antibody ABIN401183

Notes:

Passed. ABIN761396 specifically labels the CD200 in human placental tissue.

Validation Images

Staining with ABIN761396 at dilutions 1:200, 1:250, 1:300, and 1:400 (A, B, C, and D) using a 7 week elective termination of a pregnancy that tested negative for trisomies (Clark et al. (2017) Am. J. Reprod. Immunol). Optimal staining was obtained at 1:200 dilution. Anti-KLH control (E) at 1:200 was negative.

Full Methods

Primary Antibody: ABIN761396

Secondary Antibody: Bond Polymer Refine Detection kit (Leica, DS9800, lot 49232)

Full Protocol:

• Fix human placental tissue in 10% buffered formalin for 24h at RT.
• Process and embed tissue in paraffin.
• Cut paraffin blocks with a Leica CM2255 Microtome into 4µm sections.
• Affix sections to positively charged slides and air dry ON at RT.
• Deway and hydrate the slides on an automated Leica BOND Rx stainer.
• Antigen retrieval on the Leica BOND Rx automated stainer using epitope retrieval buffer 2 (Leica, AR9640, lot ER20172).
• Stain slides with primary
• • Rabbit anti-CD200 antibody (AA 45-95) antibody (antibodies-online, ABIN761396, lot 980502W),
  • rabbit anti-CD200 Receptor 1-Like (CD200R1L) (AA 150-200) antibody(antibodies-online, ABIN1715098, lot 9A13M60), or
  • rabbit anti-KLH antibody (antibodies-online, ABIN401183, lot 304770)
• diluted 1:200 in Power Vision IHC Super Blocker (Leica, PV6122). The staining protocol incorporates a modified Leica standard protocol IHC-F (which omits the post-primary step) and uses the standard times outlined in the machine protocol.
• Stain sections with Bond Polymer Refine Detection kit (Leica, DS9800, lot 49232) containing peroxidase block, post primary antibody, polymer as well as DAB chromogen and hematoxylin counterstain for times outlined in the standard protocol IHC-F.
• Remove slides from the Leica Bond Rx and then dehydrate in ethanol and clear in xylene.
• Mount slides in permount mounting medium (Fisher Scientific, SP15-500, lot 162767).
• After the slide-coverslip edges are dry, scan slides using Imagescope and photograph at 400x.

Experimental Notes:

• Several dilutions from 1:200 to 1:400 of ABIN761396 were tested and 1:200 was found to be optimal.

• The staining pattern of ABIN761396 matches our results with the rabbit anti-CD200 polyclonal antibody RB846 and observations made by others. It is also consistent with staining with the CD200R1L antibody ABIN1715098).

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-CD200 antibody (ABIN761396)

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 CD200

Target: CD200

Alternative Name: Cd200

Background:

Synonyms: MRC, MOX1, MOX2, OX-2, OX-2 membrane glycoprotein, CD200, My033

Background: Costimulates T-cell proliferation. May regulate myeloid cell activity in a variety of tissues.

Gene ID: 4345

UniProt: P41217