Recombinant Protein A/G, AF647 conjugate

Cat. No.: rp303278

GRADE & PURITY BioReagent, Azide Free, Suitable for Immunofluorescence(IF), 1.0 mg/mL

Accession #

P19909,P02976

Protein Tag

No tag

Expression system

E. coli

Overview

  Aladdin offers recombinant protein A/G conjugated to our LumiDye™ AF Dyes (Table 1).

Table 1. Spectral characteristics of protein A/G conjugates.

Cat.No.	Conjugate	λmax (nm)	Em (nm
rp303272	HRP	NA	NA
rp303273	FITC	498	517
rp303275	AF488	490	525
rp303276	AF555	553	568
rp303277	AF594	590	618
rp303278	AF647	650	668
rp302416	Cy3	554	566

Aladdin recombinant Protein A/G is a genetically engineered protein containing 5 IgG-binding regions of protein A and 2 of protein G. Cell wall binding region, cell membrane binding region and albumin binding region have been removed from the recombinant A/G-Cys to ensure the maximum specific IgG binding. The recombinant Protein A/G is ideal for making affinity gel to purification of polyclonal or monoclonal IgG antibodies. Protein A/G binds to various human, mouse and rat IgG subclasses (e.g., human IgG1, IgG2, IgG3, IgG4; mouse IgG2a, IgG2b, IgG3; rat IgG2a, IgG2c). It also binds to total IgG from cow, goat, sheep, house, rabbit, guinea pig, pig, dog and cat.

We have detected the binding data of different IgG subtypes with recombinant protein A/G, as shown in the Graph1.

Graph 1. Protein A/G affinity to selected species.

  Recombinant Protein A/G was conjugated with LumiDye™ AF647 under optimal conditions and subsequently purified to remove unconjugated free dyes. This conjugate can be used for immunoglobulin detection in Flow Cytometry and IF and for assay development. Red Fluorescence. Excitation/Emission wavelength= 650 nm/668 nm.

Application：

IF/ICC: 1:500

Note: Because staining protocols vary with application, the appropriate dilution of the protein A conjugates should be determined empirically.

Specifications

Product Name

Recombinant Protein A/G, AF647 conjugate

Synonyms

Protein A/G- AF647 | Recombinant Protein A/G- AF647 | AF647-Recombinant Protein A/G | Recombinant protein A/G, AF647 conjugated | reProtein A/G- AF647

Grade

Azide Free, BioReagent, Suitable for Immunofluorescence(IF)

Specifications & Purity

Azide Free, BioReagent, Suitable for Immunofluorescence(IF), 1.0 mg/mL

Biochemical and Physiological Mechanisms

Plays a role in the inhibition of the host innate and adaptive immune responses. Possesses five immunoglobulin-binding domains that capture both the fragment crystallizable region (Fc region) and the Fab region (part of Ig that identifies antigen) of immu

Amino Acids

35-330 aa & 373-497 aa

Sequence

NAAQHDEAQQNAFYQVLNMPNLNADQRNGFIQSLKDDPSQSANVLGEAQKLNDSQAPKADAQQNNFNKDQQSAFYEILNMPNLNEAQRNGFIQSLKDDPSQSTNVLGEAKKLNESQAPKADNNFNKEQQNAFYEILNMPNLNEEQRNGFIQSLKDDPSQSANLLSEAKKLNESQAPKADNKFNKEQQNAFYEILHLPNLNEEQRNGFIQSLKDDPSQSANLLAEAKKLNDAQAPKADNKFNKEQQNAFYEILHLPNLTEEQRNGFIQSLKDDPSVSKEILAEAKKLNDAQAPKEEDSLEGSGSGTYKLILNGKTLKGETTTEAVDAATAEKVFKQYANDNGVDGEWTYDDATKTFTVTEKPEVIDASELTPAVTTYKLVINGKTLKGETTTKAVDAETAEKAFKQYANDNGVDGVWTYDDATKTFTVTE

Conjugation

AF647

Excitation (nm)

Red Laser(633nm)

Emission(nm)

668nm

Ex/Em(nm)

Accession #

P19909, P02976

Molecule Type

Protein

Storage and Shipping

Concentration

1.0 mg/mL

Storage

Store at 2-8°C,Protected from light

Shipped In

Wet ice

Stability And Storage

The solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light.

Images

IF/ICC: Recombinant Protein A/G, AF647 conjugate (rp303278)

Recombinant Protein A/G, AF647 conjugate (rp303278) – IF/ICC 

IF analysis of EpCAM (red) in HT-29 cells. The cells were fixed with 4% Paraformaldehyde Fix Solution (P395744) for 15 minutes, and blocked with 5% BSA for 1 hour at room temperature. Cells were stained with Citatuzumab (anti-EpCAM) (Ab209902; Isotype: Human IgG1) at 1/1000 dilution in blocking buffer overnight at 4°C, and then incubated with Recombinant Protein A/G, AF647 conjugate (rp303278) at 1/500 dilution for 1 hour at room temperature in the dark (red). PBS instead of the primary antibody was used as the secondary antibody only control. Cells were counterstained with Hoechst 33342 (B113845) (blue) at 1/1000 dilution for 5 minutes. Images were taken on the confocal laser scanning microscope. .

IF/ICC: Recombinant Protein A/G, AF647 conjugate (rp303278)

Recombinant Protein A/G, AF647 conjugate (rp303278) – IF/ICC

IF analysis of EpCAM (red) in HT-29 cells. The cells were fixed with 4% Paraformaldehyde Fix Solution (P395744) for 15 minutes, and blocked with 5% BSA for 1 hour at room temperature. Cells were stained with Catumaxomab (anti-CD3&EpCAM) (Ab175489; Isotype: Mouse IgG2a & Rat IgG2b) at 1/1000 dilution in blocking buffer overnight at 4°C, and then incubated with Recombinant Protein A/G, AF647 conjugate (rp303278) at 1/500 dilution for 1 hour at room temperature in the dark (red). PBS instead of the primary antibody was used as the secondary antibody only control. Cells were counterstained with Hoechst 33342 (B113845) (blue) at 1/1000 dilution for 5 minutes. Images were taken on the confocal laser scanning microscope.

IF/ICC: Recombinant Protein A/G, AF647 conjugate (rp303278)

Recombinant Protein A/G, AF647 conjugate (rp303278) – IF/ICC 

IF analysis of EpCAM (red) in HT-29 cells. The cells were fixed with 4% Paraformaldehyde Fix Solution (P395744) for 15 minutes, and blocked with 5% BSA for 1 hour at room temperature. Cells were stained with Recombinant EpCAM Antibody (Ab101839; Isotype: Rabbit IgG) at 1/1000 dilution in blocking buffer overnight at 4°C, and then incubated with Recombinant Protein A/G, AF647 conjugate (rp303278) at 1/500 dilution for 1 hour at room temperature in the dark (red). PBS instead of the primary antibody was used as the secondary antibody only control. Cells were counterstained with Hoechst 33342 (B113845) (blue) at 1/1000 dilution for 5 minutes. Images were taken on the confocal laser scanning microscope.