Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
- Datasheet
- References (27)
- Protocols
Overview
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Product nameGoat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed
See all IgG secondary antibodies -
Host speciesGoat
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Target speciesRabbit
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Specificity
By immunoelectrophoresis and ELISA this antibody reacts specifically with rabbit IgG and with light chains common to other rabbit immunoglobulins. No antibody was detected against non-immunoglobulin serum proteins. Reduced cross-reactivity to bovine, chicken, goat, horse, human, mouse, pig and rat IgG was detected.
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Tested applicationsSuitable for: IHC-P, ICC/IF, Flow Cytmore details
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Minimal
cross-reactivity
Chicken, Cow, Goat, Horse, Human, Mouse, Pig, Rat more details -
ConjugationDyLight® 488. Ex: 493nm, Em: 518nm
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Store at +4°C.
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Storage bufferPreservative: 0.09% Sodium azide
Constituents: 0.2% BSA, PBS -
Concentration information loading... -
PurityImmunogen affinity purified
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Purification notesAntiserum was cross absorbed using bovine, chicken, horse, human, mouse, pig and rat immunosorbents to remove cross reactive antibodies. This antibody was isolated by affinity chromatography using antigen coupled to agarose beads and conjugated to DyLight® 488.
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ClonalityPolyclonal
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IsotypeIgG
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Research areas
Applications
Our Abpromise guarantee covers the use of ab96899 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| IHC-P | 1/50 - 1/500. | |
| ICC/IF | 1/50 - 1/500. | |
| Flow Cyt | 1/500. |
Images
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Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)Han et al PLoS One. 2014 Jan 23;9(1):e86539. doi: 10.1371/journal.pone.0086539. eCollection 2014. Fig 5.Effects of FZE on apoptotic ratio and apoptotic factors in RSC96 cells.
Effects of FZE on translocation of CytoC and the levels of caspase9 and caspase3. The cells were fixed with 4% paraformaldehyde for 15 minutes at 20°C, permeated with 0.3% triton prior to being blocked in 1% BSA+2% normal goat serum for 30 min at 20°C. Samples were then incubated with primary antibody overnight at 4°C in PBS containing. ab96899 diluted at 1∶200 was used as the secondary antibody. Cell nucleus were counterstained with DAPI and showed blue. Mitochondria were labeled by Mito tracker and showed red.
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Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)ICC/IF image of (ab3280) stained HeLa (Human epithelial cell line from cervix adenocarcinoma) cells.
The cells were fixed in 100% methanol (5 min) and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1 h to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab3280, 5 µg/ml) overnight at +4°C.
The secondary antibody (green) was DyLight® 488 goat anti-mouse IgG - H&L, pre-adsorbed (ab96879) used at a 1/250 dilution for 1 h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1 h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43 µM.
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Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)Emission spectra of DyLight® fluorochromes available in our catalog.
Line colors represent the approximate visible colors of the wavelength maxima. -
Flow Cytometry - Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)Overlay histogram showing A431 cells stained with unpurified ab124962 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (unpurified ab124962, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. -
Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)ab96379 staining MEK1 (phospho S298) in SK-N-SH cells treated with CNQX (ab120017), by ICC/IF. Decrease in MEK1 (phospho S298) expression correlates with increased concentration of CNQX, as described in literature.
The cells were incubated at 37°C for 24h in media containing different concentrations of ab120017 (CNQX) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab96379 (1/100 dilution) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. -
Flow Cytometry - Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)Overlay histogram showing Raji cells stained with ab52922 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab52922, 1/100) for 30 min at 22�C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22�C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Raji cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
References
This product has been referenced in:
- Pakula M et al. The Epithelial-Mesenchymal Transition Initiated by Malignant Ascites Underlies the Transmesothelial Invasion of Ovarian Cancer Cells. Int J Mol Sci 20:N/A (2019). Read more (PubMed: 30609691) »
- Xu G et al. Restraint Stress Induced Hyperpermeability and Damage of the Blood-Brain Barrier in the Amygdala of Adult Rats. Front Mol Neurosci 12:32 (2019). Read more (PubMed: 30814927) »
