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Arnt Antibody - C-terminal region (ARP30979_P050)

100 ul
$289.00
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Conjugation Options

ARP30979_P050-FITC Conjugated

ARP30979_P050-HRP Conjugated

ARP30979_P050-Biotin Conjugated

Gene Symbol:
Arnt
Official Gene Full Name:
Aryl hydrocarbon receptor nuclear translocator
NCBI Gene Id:
25242
Protein Name:
Aryl hydrocarbon receptor nuclear translocator
Swissprot Id:
P41739
Protein Accession #:
NP_036912
Nucleotide Accession #:
NM_012780
Alias Symbols:
Arnt1, Arnt
Replacement Item:
This antibody may replace item sc-17811 from Santa Cruz Biotechnology.
Description of Target:
Arnt is required for activity of the Ah (dioxin) receptor. This protein is required for the ligand-binding subunit to translocate from the cytosol to the nucleus after ligand binding. The complex then initiates transcription of genes involved in the activation of PAH procarcinogens. The heterodimer with HIF1A or EPAS1/HIF2A functions as a transcriptional regulator of the adaptive response to hypoxia.
Protein Size (# AA):
800
Molecular Weight:
88kDa
Host:
Rabbit
Clonality:
Polyclonal
Purification:
Affinity Purified
Application:
WB, IHC
Tissue Tool:
Find tissues and cell lines supported by DNA array analysis to express Arnt.
RNA Seq:
Find tissues and cell lines supported by RNA-seq analysis to express Arnt.
Immunogen:
The immunogen is a synthetic peptide corresponding to a region of Rat
Predicted Species Reactivity:
Cow, Dog, Guinea Pig, Horse, Human, Mouse, Rabbit, Rat, Sheep
Tested Species Reactivity:
Human, Rat
Predicted Homology Based on Immunogen Sequence:
Cow: 100%; Dog: 100%; Guinea Pig: 100%; Horse: 100%; Human: 100%; Mouse: 92%; Rabbit: 92%; Rat: 100%; Sheep: 100%
Complete computational species homology data:
Anti-Arnt (ARP30979_P050)
Peptide Sequence:
Synthetic peptide located within the following region: SNEQHVQPTSAQPSSQPEVFQEMLSMLGDQSNTYNNEEFPDLTMFPPFSE
Product Format:
Liquid. Purified antibody supplied in 1x PBS buffer with 0.09% (w/v) sodium azide and 2% sucrose.
Reconstitution and Storage:
For short term use, store at 2-8C up to 1 week. For long term storage, store at -20C in small aliquots to prevent freeze-thaw cycles.
Concentration:
Batch dependent within range: 100 ul at 0.5 - 1 mg/ml
Protein Interactions:
Ahr;
Blocking Peptide:
For anti-Arnt (ARP30979_P050) antibody is Catalog # AAP30979 (Previous Catalog # AAPS08305)
Datasheets/Manuals:
Printable datasheet for anti-Arnt (ARP30979_P050) antibody
Publications:

Kurlak, LO; Mistry, HD; Cindrova-Davies, T; Burton, GJ; Broughton Pipkin, F; Human placental renin-angiotensin system in normotensive and pre-eclamptic pregnancies at high altitude and after acute hypoxia-reoxygenation insult. 594, 1327-40 (2016). WB, IHC, Cow, Dog, Guinea Pig, Horse, Human, Mouse, Rabbit, Rat, Sheep 26574162

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02/01/2017 15:23
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Product Review: Arnt antibody - C-terminal region (ARP30979_P050) in Human placenta using IHC

Product Page for Arnt antibody - C-terminal region (ARP30979_P050)

Researcher: Hiten D. Mistry, King's College London
Application: IHC
Species + Tissue/Cell type: Human placenta
Primary antibody dilution: 1:200
Secondary antibody: Anti-rabbit-HRP
Secondary antibody dilution: 1:5000



Questionnaire:  
How do Aviva's reagents play a role in your experimental goals? It is one of our main antibodies of interest.
How would you rate this antibody on a scale from 1-5 (5=best) and why? 4
Would you use this antibody in future experiments? Yes
Have you used another antibody which has worked in your application? No
Do you believe the information about the reagent on Aviva's website is correct? Yes, apart from the storage conditions; this antibody is not stable for short-term 4 degree C storage as stated and should be stored at -20 degree C.
If the antibody works, do you plan to use it in future experiments or to publish your data? Why or why not? Yes
How did you store the antibody after re-suspension? -20 d C
Sample Description (please include species type and tissue/cell type): Human Placental tissue samples, paraffin-embedded slides.
Please explain fixation solution/method used (formalin, periodate-lysine-paraformaldehyde, acetone, etc.)? Formalin fixed
How many different experimental trials were conducted using the antibody sample? 6
Primary antibody dilution, incubation time and temperature: 1 in 200
Secondary antibody used, dilution, incubation time and temperature: DAKO Envision Labelled polymer HRP Anti-Rabbit (ready to use)
From your IHC/ICC images, briefly explain the colors of each stain and counterstain: DAB staining and counter stained with haemotoxylin
Did you use an antigen retrieval method? If so, please explain? Heat-induced antigen retrieval method.
What controls were used in your experiment? Negative control: IgG rabbit
Please include your detailed tissue preparation and staining procedure/protocol here: Protocol:
1. Dewax sections in xylene baths (2x5mins)
2. Immerse in absolute alcohol (IMS is cheaper) (2x5 mins)
3. Immerse in fresh 0.5% H2O2 in methanol-peroxidase block (store up to max 1wk at 4 d C) (10 mins)
4. Immerse in alcohol (2x5 mins)
5. Immerse in 70% alcohol (5 mins)
6. Wash in runnig water (5 mins)
7. Microwave for 15 mins at medium setting in Citrate buffer solution (Heat induced epitope retieval). Time depends on equipment [20 mins on high power]
8. Leave to stand for a few mins before cooling in running water [at least 15 mins total]
9. Wash in TBS (5 mins)
10. Wipe off excess TBS and draw around section with hydrophobic barrier pen pen, block with appropriate for secondary Ab normal serum (dil 1/10 in TBS/BSA) (30 mins)
11. Incubate in primary Ab (dil in BS/BSA) overnight at 4 d C, then 30-60 mins at room temp.
12. Wash in TBS (2x6 mins)
13. Incubate with HRP-labelled polymer (secondary Ab) at room temp, neat if part of kit or concentration as assessed in a validation run (30 mins)
14. Wash in TBS (2x5 mins)
15. In cubate in DAB chromogen [make up in separate tube at 1ml of buffer + 20ul DAB] (1x10 mins)
16. Wash in running water for a few minutes
17. Counter stain in Harris' Haematoxylin (1x12 secs)
18. Wash in running water
19. Blue in Scott's Tap water. Look at slide under microscope (5 secs)
20. Wash in running water
21. If too blue, dip in acid alcohol.
22. Wash in running water
23. Dip in Scott's Tap water (5 secs)
24. Wash in running water
25. Transport in water/TBS
26. Immerse in 70% alcohol (5 mins)
27. Immerse in 100% alcohol (2x5 mins)
28. Clear in xylene (III and iv) (2x5 mins)
29. Mount in DPX
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