RFP 抗体 (Red Fluorescent Protein) (AA 234)

Details for Product anti-RFP Antibody No. ABIN1043867, 供应商: Log in to see
抗原
抗原表位
AA 234
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适用
Discosoma
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宿主

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克隆类型
多克隆
标记
This RFP antibody is un-conjugated
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应用范围
Fluorescence Microscopy (FM), Immunohistochemistry (IHC), ELISA, Western Blotting (WB)
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Supplier
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Available images

'独立验证'标志
抗原 RFP
Lot Number 34945
Method validated Immunofluorescence
Positive Control CX3CL1mcherry+ murine thymus
Negative Control CX3CL1 WT murine thymus
Notes

Passed. The RFP antibody ABIN1043867 specifically labels RFP+ cells in CX3CL1mcherry+ murine thymus in immunofluorescence, consistent with the expression pattern of CX3CL1mcherry.

Primary Antibody ABIN1043867
Secondary Antibody donkey Fab'2 anti-rabbit A647 conjugated antibody (Jackson Immunoresearch, 711-606-152, lot 128806)
实验流程
  • Harvest thymus from 7 weeks old mice in 1X Dulbecco's phosphate buffered saline (DPBS) (Gibco Life Technologies, 14200-067).
  • Fix mouse thymus in antigen fix (Diapath, P0014) for 2h at 4?C.
  • Wash tissue in 0.1M pH7.4 phosphate buffer at for 1h at 4?C.
  • Dehydrate tissue in 30% sucrose solution at 4?C ON.
  • Snap freeze tissue in Tissue Freezing Medium (ElectroMicroscopy Science, 72592-C) at -80?C.
  • Cut blocks into 25?m sections using a cryostat (Leica, CM3050 S).
  • Transfer sections to a slide.
  • Create a hydrophobic barrier on the slide around sections with Dako pen (Dako, S2002, lot 00081640).
  • Place slide in a humidified chamber and rehydrate sections in 0.1 M TrisHCl pH7.4 for 10min at RT.
  • Gently remove buffer by tapping slide.
  • Permeabilize tissue in 0.1M TrisHCl pH7.4 containing 2% Triton X-100 (Sigma, lot 015K0039) and 0.5% BSA, for 20min at RT.
  • Incubate sections with primary RFP antibody (Red Fluorescent Protein) (AA 234) (antibodies-online, ABIN4368323, lot 1602) diluted 1:1000 0.1M TrisHCl pH7.4 containing 2% Triton X-100 (Sigma, batch 015K0039) and 0.5% BSA for 2h at RT.
  • Incubate a no primary antibody negative control in parallel in 0.1M TrisHCl pH7.4 containing 2% Triton X-100 (Sigma, batch 015K0039) and 0.5% BSA.
  • Wash slides 1x 5min in 0.1M TrisHCl pH7.4.
  • Incubate sections with secondary donkey Fab'2 anti-rabbit AF647 conjugated antibody (Jackson Immunoresearch, 711-606-152, lot 128806) diluted 1:300 in 0.1M TrisHCl pH7.4 containing 2% Triton X-100 (Sigma, batch 015K0039) and 0.5% BSA for 2h at RT.
  • Wash slides 1x 5min in 0.1M TrisHCl pH7.4.
  • Add approximately 10?L of Slowfade Gold antifade reagent (ThermoFisher Scientific, S36937, lot 1226836) for each section and mount cover slip.
  • Image acquisition on an LSM 880 (Zeiss), 20x magnification, 1000 resolution.
Validation Images
Immunofluorescence validation image for anti-Red Fluorescent Protein (RFP) (AA 234) antibody (ABIN1043867)

CX3CL1 positive cells in CX3CL1mcherry murine thymus (A, mCherry) were stained wit...

免疫原 The immunogen is a Red Fluorescent Protein (RFP) fusion protein corresponding to the full length amino acid sequence (234aa) derived from the mushroom polyp coral Discosoma.
Immunogen Type: Recombinant Protein
亚型 IgG
特异性 This product was prepared from monospecific antiserum by imunoaffinity chromatography using Red Fluorescent Protein (Discosoma) coupled to agarose beads followed by solid phase adsorption(s) to remove any unwanted reactivities. Expect reactivity against RFP and its variants: mCherry, tdTomato, mBanana, mOrange, mPlum, mOrange and mStrawberry. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Rabbit Serum and purified and partially purified Red Fluorescent Protein (Discosoma). No reaction was observed against Human, Mouse or Rat serum proteins. ELISA was used to confirm specificity at less than 0.1% of target signal.
产品特性 Antibodies to RFP (Discosoma spp.) are intended for use in immunological assays including ELISA, western blotting, fluorometry and fluorescence activated cell sorting (FACS).
过滤 Sterile filtered
别名 RFP
背景 Antibodies to RFP (Discosoma spp.) are intended for use in immunological assays including ELISA, western blotting, fluorometry and fluorescence activated cell sorting (FACS).
Synonyms: DsRed, rDsRed, Discosoma sp. Red Fluorescent Protein, Red fluorescent protein drFP583
研究领域 Tags/Labels
应用备注 Suggested dilutions:
ELISA : 1:90000
IF Microscopy : 1:200-1:2000
Western Blot : 1:2000 - 1:10000
Immunochemistry: 1:200-1:2000
Other Dilution: User Optimized
说明

Gene Name: DsRed

限制 仅限研究用
状态 Liquid
浓度 1.0 mg/mL
缓冲液 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
储存液 Sodium azide
注意事项 This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
储存条件 4 °C/-20 °C
储存方法 Store vial at 4 °C prior to restoration. For extended storage aliquot contents and freeze at -20 °C or below. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4 °C as an undiluted liquid. Dilute only prior to immediate use. Expiration date is one (1) year from date of opening.
有效期 12 months
厂商提供的图像
Western Blotting (WB) image for anti-RFP 抗体 (Red Fluorescent Protein) (AA 234) (ABIN1043867) Western blot of RFP recombinant protein detected with polyclonal anti-RFP antibody. L...
有引用在: Viganò, Möbius, Götz, Dimou: "Transplantation reveals regional differences in oligodendrocyte differentiation in the adult brain." in: Nature neuroscience, Vol. 16, Issue 10, pp. 1370-2, 2013 (PubMed). (1:500).

Gross, Baird, Hoffman, Baldridge, Tsien: "The structure of the chromophore within DsRed, a red fluorescent protein from coral." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 97, Issue 22, pp. 11990-5, 2000 (PubMed).