Goatanti-RabbitIgG(Heavy&LightChain)Antibody(FITC) 抗体网

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产品名称：Goatanti-RabbitIgG(Heavy&LightChain)Antibody(FITC)
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产品厂商：BioLeaf
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简单介绍：

Goatanti-RabbitIgG(Heavy&LightChain)Antibody(FITC)

详情介绍：

	Successfully validated (Immunofluorescence) by Okeanos Research Laboratory, Department of Biological Sciences, Clemson University No. #100071 Date 09/14/2016
Antigen	Rabbit IgG (Heavy & Light Chain)
Lot Number	611-1202
Method validated	Immunofluorescence
Positive Control	Lab stock CBD-SNAP antibody
Negative Control	No SNAP-tag antibody
Notes	We validate the specificity of the secondary goat anti-rabbit IgG (heavy & light chain) antibody (FITC) ABIN101988 for rabbit IgG antibody.

Full Methods
Full Methods
Primary Antibody	ABIN1573927
Secondary Antibody	ABIN101988
Protocol	Oyster visceral mass tissue is dissected and fixed in 4% paraformaldehyde in seawater overnight. Serial dehydration process using an automated ASP300S Enclosed Tissue Processor (Leica Biosystems) as follows: 70% ethanol for 45min 90% ethanol for 45min 90% ethanol for 45min 100% ethanol twice for 45min xylene twice for 45min paraffin wax at 58°C 3 times for 30 min Tissue is mounted in a paraffin block and hardened overnight before. 8µm tissue sections are retrieved from the block and collected on circular glass cover slips. Heat cover slips at 60°C for 1h. Deparaffination and rehydration: Xylene twice for 15min 100% ethanol twice for 10min 95% ethanol for 10 min 85% ethanol for 10 min 70% ethanol for 10 min 50% ethanol for 10 min 30% ethanol for 10 min distilled water for 10 min PBS for 10 min Wash tissue sections with PBS with 0.05% triton X twice for 30min. Permeabilize in PBS with 0.05% triton X overnight. Treatment of the tissue sections with 1mg/mL sodium borohydride in PBS three times for 5min to reduce autofluorescence. Wash sections in PBS 3 times for 15 min for at RT. Block sections in PBST with 1% BSA for 2 hours at RT. Incubate sections with CBD-SNAP antibody (lab stock) diluted 1:200 in PBST with 1% BSA overnight at 4°C to detect the location of chitin. Wash sections in PBS 3 times for 15min with PBS at RT. Additionally, incubate the CBD-SNAP and SNAP-tag double-stained sections with rabbit anti-SNAP antibody (antibodies-online, ABIN1573927, lot 13D000621) diluted 1:200 in PBST with 1% BSA overnight at 4°C. Wash sections in PBS 3 times for 15min with PBS at RT. Incubate sections with the secondary goat anti-rabbit IgG (heavy & light chain) antibody (FITC) (antibodies-online, ABIN101988, lot 611-1202) diluted 1:400 in PBST with 1% BSA for 2h at °C. Wash sections in PBS three times for 15min at RT. Counterstain with 0.1µg/mL DAPI in PBS for 15min at RT. Wash sections in PBS three times for 15min at RT. Mount sections on a microscopic slide using 50% glycerol in PBS. Seal cover slips with nail polish. Confocal imaging on Leica SPE. Visualization of the data performed on LAS 3D software.
Experimental Notes	To validate the specificity of the anti-rabbit FITC secondary antibody ABIN101988, 8µm paraffin sections of oyster visceral mass were observed in this study. We compared the fluorescence signals with immunofluorescence study. The negative control specimen was always compared with the test specimen or the positive control specimen on the same day, using the same laser power, gain, offset, accumulation/averaging settings on the Leica SPE confocal microscope. Visualization of the data was performed on LAS 3D software, with the same visualization setting to compare signal brightness. We found that the samples treated with anti-rabbit FITC secondary antibody ABIN101988 had similar fluorescence signals as the positive control Anti Rabbit Alexa 488. Excitation at the same laser wavelength and power did not generate fluorescence in the negative control section, when anti-rabbit FITC secondary antibody was applied in the absence of rabbit produced anti-SNAP antibody.

Images
Images
Validation Images	Immunofluorescence images of oyster visceral mass tissue, with the specificity of Ant... Immunofluorescence images of oyster visceral mass tissue, with the specificity of Anti-Rabbit Alexa 488 (top row) and Anti-Rabbit FITC (bottom row) compared. Unstained samples (left column) were compared to test samples (right column) to visualize degree of auto-fluorescence. The absence of anti-SNAP served as the negative control (middle column), it has led to the absence of fluorescence at the same imaging and visualization setting for the green channels, implying the goat anti-rabbit IgG (Heavy & Light Chain) antibody FITC conjugate ABIN101988 is specific to Rabbit IgG antigen. Immunofluorescence images of oyster visceral mass tissue, with the specificity of Anti-Rabbit Alexa 488 (top row) and Anti-Rabbit FITC (bottom row) compared. Unstained samples (left column) were compared to test samples (right column) to visualize degree of auto-fluorescence. The absence of anti-SNAP served as the negative control (middle column), it has led to the absence of fluorescence at the same imaging and visualization setting for the green channels, implying the goat anti-rabbit IgG (Heavy & Light Chain) antibody FITC conjugate ABIN101988 is specific to Rabbit IgG antigen.

Product details
Product details
Immunogen	Rabbit IgG whole molecule
Isotype	IgG
Specificity	IgG (H&L)
Characteristics	Concentration Definition: by UV absorbance at 280 nm

Target details
Target details
Research Area	Immunology, Secondary Antibodies

Application Details
Application Details
Application Notes	This product is designed for immunofluorescence microscopy, fluorescence based plate assays (FLISA) and fluorescent western blotting. This product is also suitable for multiplex analysis, including multicolor imaging, utilizing various commercial platforms.
Comment	Excitation/Emission wavelength: 494 nm/514 nm
Restrictions	For Research Use only

Handling
Handling
Format	Lyophilized
Reconstitution	Restore with deionized water (or equivalent)
Concentration	2.0 mg/mL
Buffer	0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Preservative	Sodium azide
Precaution of Use	WARNING: Reagents contain sodium azide. Sodium azide is very toxic if ingested or inhaled. Avoid contact with skin, eyes, or clothing. Wear eye or face protection when handling. If skin or eye contact occurs, wash with copious amounts of water. If ingested or inhaled, contact a physician immediately. Sodium azide yields toxic hydrazoic acid under acidic conditions. Dilute azide-containing compounds in running water before discarding to avoid accumulation of potentially explosive deposits in lead or copper plumbing.
Handling Advice	Product is photosensitive and should be protected from light.

Images
Images
Supplier Images	FITC (fluorescein) and HRP (horse radish peroxidase) conjugated secondary antibody wa... FITC (fluorescein) and HRP (horse radish peroxidase) conjugated secondary antibody was used to detect nanogram – picogram levels of rabbit IgG by dot blot on nitrocellulose membrane. 4 ul each of serial 1 in 4 dilutions of rabbit IgG were dotted on nitrocellulose and allowed to dry. Membrane was blcoked in 3% BSA for 10 minutes dried for later use and rewetted with MB-070. Blot was incubated in fluorescein conjugated goat anti rabbit 611-1202 lot 25176 1:10,000 and HRP conjugated goat anti Rabbit (611-1302 lot 25406 1:10,000, dried and: A. Blot was imaged on the BioRad VersaDoc with filter settings appropriate for Fluorescein/DyLight 488 B. Blot was rewetted with TBS, incubated with FEMTOMAX chemiluminescent substrate for 1-3 minutes and imaged for 60sec on the BioRad VersaDoc Imaging System C. Blot was rinsed with TBS and DIH2O, incubated for 5 minutes with TMB Substrate for Western Blot MaxTag (1 ml of TMBM-102 + ~9 ml of TMBM-101), dried overnight and imaged using a conventional flatbed scanner FITC (fluorescein) and HRP (horse radish peroxidase) conjugated secondary antibody was used to detect nanogram – picogram levels of rabbit IgG by dot blot on nitrocellulose membrane. 4 ul each of serial 1 in 4 dilutions of rabbit IgG were dotted on nitrocellulose and allowed to dry. Membrane was blcoked in 3% BSA for 10 minutes dried for later use and rewetted with MB-070. Blot was incubated in fluorescein conjugated goat anti rabbit 611-1202 lot 25176 1:10,000 and HRP conjugated goat anti Rabbit (611-1302 lot 25406 1:10,000, dried and: A. Blot was imaged on the BioRad VersaDoc with filter settings appropriate for Fluorescein/DyLight 488 B. Blot was rewetted with TBS, incubated with FEMTOMAX chemiluminescent substrate for 1-3 minutes and imaged for 60sec on the BioRad VersaDoc Imaging System C. Blot was rinsed with TBS and DIH2O, incubated for 5 minutes with TMB Substrate for Western Blot MaxTag (1 ml of TMBM-102 + ~9 ml of TMBM-101), dried overnight and imaged using a conventional flatbed scanner

References
References
Background publications	Blakeslee, Baines: "Immunofluorescence using dichlorotriazinylaminofluorescein (DTAF). I. Preparation and fractionation of labelled IgG." in: Journal of immunological methods, Vol. 13, Issue 3-4, pp. 305-20, 1977 (PubMed).