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RFP 抗体

Name: RFP 抗体
SKU: ABIN967350
Availability: OutOfStock
Rating: 5 (1 reviews)

RFP 适用: Discosoma WB 宿主: 小鼠 Monoclonal E64-1077 unconjugated

Western Blotting validation image for anti-Red Fluorescent Protein (RFP) antibody (ABIN967350)

2 images

(1 reference)

(1 validation)

产品编号 ABIN967350

发货至: 中国

Datasheet as PDF

抗原 See all RFP 抗体

RFP (Red Fluorescent Protein (RFP))

适用

Discosoma
宿主
23

13

9

5

3
小鼠
Compatible Secondaries
克隆类型
30

23
单克隆
标记
19

5

3

2

2

2

2

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1
This RFP antibody is un-conjugated
应用范围
27

21

15

12

10

7

6

6

2

1

1

1
Western Blotting (WB)

品牌

BD Pharmingen™

产品特性

1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Please refer to us for technical protocols.
3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
4. Source of all serum proteins is from USDA inspected abattoirs located in the United States.

纯化方法

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

免疫原

DsRed1 recombinant protein

克隆位点

E64-1077

亚型

IgG1

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应用备注

Applications include western blot analysis (0.25-1.0 µg/ml). DsRed2 control lysate is provided as a positive control (component 51-16616N) and approximately 10 µl is recommended to visualize DsRed by western blot analysis.

说明

Related Products: ABIN967389

限制

仅限研究用
生效 #101231 (Western Blotting)

by

Institut für Virologie, Medizinische Hochschule Hannover

No.

#101231

日期

2017.06.19

抗原

DsRed

Lot Number

6223801

Method validated

Western Blotting

Positive Control

Human foreskin fibroblasts infected with DsRed expressing virus

Negative Control

Cells infected with EGFP expressing virus

Notes

Passed. ABIN967350 specifically recognizes DsRed in extracts of cells expressing DsRed and GFP.

Validation Images

Immunoblot of DsRed or EGFP expressing human fibroblast lysates using ABIN967350. Cells were infected with EGFP-expressing virus (V1-gfp), DsRed-expressing viruses (V2-/V3-DsRed), or co-infected as indicated. The lane on the far right contains a sample from non-infected cells.
Full Methods
Primary Antibody

ABIN967350

Secondary Antibody

goat anti-mouse antibody (Pierce, 32230, lot QC212446)

Full Protocol

Grow fibroblasts in Dulbecco’s modified Eagle’s medium (Biochrom, F-0435, lot 0315F) supplemented with 10% fetal calf serum (FBS Good Forte, PAN Biotech, P40-47500, lot P130912)and 100U/ml penicillin, 100µg/ml streptomycin sulfate (PAN Biotech, P06-19100, lot 7530716) at 37°C and 5% CO₂ in 2ml on a tissue culture 6-well plate to 90% confluency.
Infect cells at an MOI of 3 with
an EGFP-expressing virus V1-gfp,
a DsRed expressing viruse V2-DsRed,
a DsRed expressing viruse V3-DsRed,
virus V1-gfp and V2-DsRed, or
virus V1-gfp and V3-DsRed
Grow cells for 4d at 37°C and 5% CO₂.
Collect 8.5x10⁴ cells directly in 200µl of1x Laemmli SDS sample buffer, and boil them for 5min at 95°C.
Separate the equivalent of 5000 cells of each sample on an Any kD SDS-PAGE gel (Bio-Rad, 456-9036, control 400100025) in a Bio-Rad Mini-Protean gel electrophoresis chamber for 45min at 250V and 21°C.
Transfer proteins onto nitrocellulose membrane (GE Healthcare, 10600003, lot G7724136) with a TE77X semi-dry blotting system (SERVA) for 30min at 0.5A.
Block the membrane using Roti-Block solution (Carl Roth, A151.1, lot 382190165) for 1h at RT.
Incubate the membrane with primary
mouse anti-Red Fluorescent Protein (DsRed) antibody (antibodies-online, ABIN967350, lot 6223801)
rabbit anti-GFP (D5.1) XP (Cell Signalling, 2956, lot 4) antibody
diluted 1:1000 in Roti-Block solution ON at RT.
Wash the membrane 4x for 5min in PBS + 0.1% Tween20 (PBST).
Incubate the membrane with secondary HRP-conjugated
goat anti-mouseantibody (Pierce, 32230, lot QC212446)
goat anti-rabbitantibody (Pierce, 32260, lot QH219992)
diluted 1:2000 in PBST for 1h at RT.
Wash the membrane 4x for 5min in PBST.
Visualize protein bands using SuperSignal West Femto Maximum Sensitivity Substrate (Thermo Scientific, 34096, lot RH236205) on a LAS 3000 imager (Fuji).
Strip the membrane using Restore PLUS Western blot stripping buffer (Thermo Scientific, 46428, lot PA196499) for 20min at RT.
Wash the membrane 4x for 5min in PBST, and block for 1h at RT with Roti-Block solution.
Incubate the membrane with primary rabbit anti-GAPDH antibody (Cell signalling, 2118, lot 10) diluted 1:2000 in PBST for 1h at RT.
Wash the membrane 4x for 5min in PBST.
Incubate the membrane with secondary goat anti-rabbit antibody (PIERCE, 32260, lot QH219992) diluted 1:2000 in PBST for 1h at RT.
Wash the membrane 4x for 5min in PBST.
Visualize protein bands using SuperSignal West Femto Maximum Sensitivity Substrate (Thermo Scientific, 34096, lot RH236205) on a LAS 3000 imager (Fuji).

Experimental Notes

The Red Fluorescent Protein (DsRed) antibody ABIN967350 reveals a protein of the expected molecular weight (31kDa) in lysates of cells infected with DsRed or with DsRed and EGFP expressing virus. A slightly lower MW band is visible in the lysates of EGFP (expected MW 28kDa) expressing cells. This might reflect some cross-reactivity of the DsRed antibody with the structurally similar EGFP. However, the overall selectivity of ABIN967350 towards DsRed is very good.
状态

Liquid

浓度

0.25 mg/mL

缓冲液

Aqueous buffered solution containing BSA, glycerol, and ≤0.09 % sodium azide.

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

-20 °C

储存方法

Store undiluted at -20°C.
Wall, Socolich, Ranganathan: "The structural basis for red fluorescence in the tetrameric GFP homolog DsRed." in: Nature structural biology, Vol. 7, Issue 12, pp. 1133-8, (2000) (PubMed).
抗原

RFP (Red Fluorescent Protein (RFP))

别名

DsRed Fluorescent Protein (RFP 产品)

物质类

Chemical

背景

DsRed is naturally occurring fluorescent protein from a coral of the genus Discosoma. DsRed, like green fluorescent protein (GFP) can be introduced into either cultured cells or in transgenic animals to study the localization of a specific protein. It can also be used in fusion constructs with a protein of interest to examine protein-protein interactions or as a general marker. The crystal structure of DsRed has been solved and shown that DsRed monomers exhibit similar topology to GFP, but have additional chemical modifications which account for differences in the spectra, when compared to GFP. DsRed has a longer emission/absorption wavelength than GFP. DsRed migrates at 31 kDa in SDS/PAGE. The antibody recognizes three forms of DsRed, 1, 2 and pTimer. A recombinant protein representing DsRed1 was used as the immunogen.

分子量

31 kDa