Anti-Aly/Ref antibody [EPR17942] (ab202894)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR17942] to Aly/Ref
- Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IP
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-Aly/Ref antibody [EPR17942]
See all Aly/Ref primary antibodies -
Description
Rabbit monoclonal [EPR17942] to Aly/Ref -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species Flow Cyt HumanICC/IF RatHumanIHC-P MouseRatHumanIP HumanWB MouseRatHuman -
Immunogen
This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: PC-3, HeLa, HepG2, HEK293 whole cell lysates; Human fetal brain, fetal heart and fetal kidney lysates; Mouse brain and heart lysates; rat brain and heart lysates. IHC-P: Human cervix carcinoma, Mouse cerebral cortex and rat liver tissues. ICC/IF: HeLa and C6 cells. Flow Cyt: HeLa cells. IP: HepG2 whole cell lysate.
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General notes
This product was previously labelled as Aly
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
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. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR17942 -
Isotype
IgG -
Research areas
Images
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All lanes : Anti-Aly/Ref antibody [EPR17942] (ab202894) at 1/10000 dilution
Lane 1 : PC-3 (Human prostate cancer cell line) whole cell lysate
Lane 2 : HepG2 (Human liver hepatocellular carcinoma) whole cell lysate
Lane 3 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
Lane 4 : HEK293 (Human epithelial cells from embryonic kidney) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution
Predicted band size: 27 kDa
Observed band size: 27 kDa
Exposure time: 5 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-Aly/Ref antibody [EPR17942] (ab202894) at 1/2000 dilution
Lane 1 : Human fetal brain lysate
Lane 2 : Human fetal heart lysate
Lane 3 : Human fetal kidney lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 27 kDa
Observed band size: 27 kDa
Exposure time: 15 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-Aly/Ref antibody [EPR17942] (ab202894) at 1/2000 dilution
Lane 1 : Mouse brain lysate
Lane 2 : Mouse heart lysate
Lane 3 : Rat brain lysate
Lane 4 : Rat heart lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution
Predicted band size: 27 kDa
Observed band size: 27 kDa
Exposure time: 15 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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Immunohistochemical analysis of paraffin-embedded Human cervix carcinoma tissue labeling Aly/Ref with ab202894 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear and weakly cytoplasmic staining on Human cervix carcinoma tissue is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded mouse cerebral cortex tissue labeling Aly/Ref with ab202894 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear and weakly cytoplasmic staining on mouse cerebral cortex tissue is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded rat liver tissue labeling Aly/Ref with ab202894 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear and weakly cytoplasmic staining on rat liver tissue is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Aly/Ref with ab202894 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HeLa cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: ab202894 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution. -
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized C6 (Rat glial tumor cells) cells labeling Aly/Ref with ab202894 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear and weakly cytoplasmic staining on C6 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: ab202894 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution. -
Flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Aly/Ref with ab202894 at 1/150 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/500 dilution was used as the secondary antibody.
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Aly/Ref was immunoprecipitated from 1mg of HepG2 (Human liver hepatocellular carcinoma) whole cell lysate with ab202894 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab202894 at 1/5000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.
Lane 1: HepG2 whole cell lysate 10 µg (Input). Lane 2: ab202894 IP in HepG2 whole cell lysate. Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab202894 in HepG2 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 8 seconds.
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