Anti-RIG-I/DDX58 antibody (ab45428)
Key features and details
- Rabbit polyclonal to RIG-I/DDX58
- Suitable for: ICC/IF, IHC-P, WB
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
Overview
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Product name
Anti-RIG-I/DDX58 antibody
See all RIG-I/DDX58 primary antibodies -
Description
Rabbit polyclonal to RIG-I/DDX58 -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, IHC-P, WBmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Full length protein (GST-tag) corresponding to Human RIG-I/DDX58.
Database link: O95786 -
Positive control
- ICC/IF: MCF7 and HeLa cells. IHC-P: Human, rat and mouse testis. WB: Rat heart tissue and A20 cell lysates.
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General notes
This product is not recommended for WB using human samples as it is not able to detect human DDX58 in this application. We recommend ab180675 as a knockout validated product for testing human samples in WB.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.
One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.
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Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.2
Preservative: 0.02% Sodium azide
Constituent: PBS -
Concentration information loading... -
Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RIG-I/DDX58 antibody (ab45428)
Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-RIG1/DDX58 antibody (ab45428) at 1 ug/ml. Tissue was fixed with formaldehyde and blocked with 5% BSA for 1 h at RT; antigen retrieval was by heat mediation (microwave for 20 minutes) with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. Streptavidin-HRP Kit (Rabbit IgG) at 1/200 was used as secondary at room temperature for 1 hour. Counter stained with Hematoxylin.
Endogenous peroxidase blocking: 3% H202 at room temperature for 10 minutes.
Washing: PBS, 2 x 5 minutes.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RIG-I/DDX58 antibody (ab45428)Immunofluorescent analysis of 10% formalin-fixed (24 hours) mouse testis labeling RIG-1/DDX58 with ab45428 at 20 ug/mL, followed by Rabbit IgG antibody (Alexa Fluor 488) at 1/2000 dilution (green) and DAPI staining (blue).
Blocking condition: 5% BSA, room temperature for 1 hour.
Primary antibody incubation: 4°C overnight.
Secondary antibody incubation: room temperature for 1 hour.
Washing condition: PBS, 3 x 3 minutes
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Immunocytochemistry/ Immunofluorescence - Anti-RIG-I/DDX58 antibody (ab45428)Immunofluorescent analysis of 4% paraformaldehyde-fixed (room temperature for 10 minutes) MCF7 cells labeling RIG-1/DDX58 with ab45428 at 20 ug/mL, followed by Rabbit IgG antibody (Alexa Fluor 488) at 1/2000 dilution (green) and DAPI staining (blue).
Permeabilization condition: 0.1% Triton X-100 in PBS, room temperature, 5 minutes.
Blocking condition: 5% BSA, room temperature for 1 hour.
Primary antibody incubation: 4°C overnight (1/50 dilution).
Secondary antibody incubation: room temperature for 1 hour.
Washing condition: PBS, 3 x 3 minutes
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RIG-I/DDX58 antibody (ab45428)Immunofluorescent analysis of 10% formalin-fixed (24 hours) rat testis labeling RIG-1/DDX58 with ab45428 at 20 ug/mL, followed by Rabbit IgG antibody (Alexa Fluor 488) at 1/2000 dilution (green) and DAPI staining (blue).
Blocking condition: 5% BSA, room temperature for 1 hour.
Primary antibody incubation: 4°C overnight.
Secondary antibody incubation: room temperature for 1 hour.
Washing condition: PBS, 3 x 3 minutes
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RIG-I/DDX58 antibody (ab45428)Immunofluorescent analysis of 10% formalin-fixed (24 hours) human testis labeling RIG-1/DDX58 with ab45428 at 20 ug/mL, followed by Rabbit IgG antibody (Alexa Fluor 488) at 1/2000 dilution (green) and DAPI staining (blue).
Blocking condition: 5% BSA, room temperature for 1 hour.
Primary antibody incubation: 4°C overnight.
Secondary antibody incubation: room temperature for 1 hour.
Washing condition: PBS, 3 x 3 minutes
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Western blot - Anti-RIG-I/DDX58 antibody (ab45428)All lanes : Anti-RIG-I/DDX58 antibody (ab45428) at 2 µg/ml
Lane 1 : A20 cell lysate
Lane 2 : Rat heart tissue lysate
Predicted band size: 107 kDa
Observed band size: 120 kDa why is the actual band size different from the predicted?The band of interest is at 120 kDa. Bands below 120 kDa are non-specific.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RIG-I/DDX58 antibody (ab45428)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat heart tissue labelling RIG-I/DDX58 with ab45428 at a dilution of 10µg/ml.
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Immunocytochemistry/ Immunofluorescence - Anti-RIG-I/DDX58 antibody (ab45428)ICC/IF image of ab45428 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab45428, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
