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NOS2 Antibody - N-terminal region (ARP63000_P050)

100 ul
$289.00
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Conjugation Options

ARP63000_P050-FITC Conjugated

ARP63000_P050-HRP Conjugated

ARP63000_P050-Biotin Conjugated

Gene Symbol:
NOS2
Official Gene Full Name:
Nitric oxide synthase 2, inducible
NCBI Gene Id:
4843
Protein Name:
Nitric oxide synthase, inducible
Swissprot Id:
P35228
Protein Accession #:
EAW51048
Nucleotide Accession #:
NM_000625
Alias Symbols:
HEP-NOS, INOS, NOS, NOS2A
Replacement Item:
This antibody may replace item sc-49055 from Santa Cruz Biotechnology.
Description of Target:
Nitric oxide is a reactive free radical which acts as a biologic mediator in several processes, including neurotransmission and antimicrobial and antitumoral activities. This gene encodes a nitric oxide synthase which is expressed in liver and is inducible by a combination of lipopolysaccharide and certain cytokines. Three related pseudogenes are located within the Smith-Magenis syndrome region on chromosome 17.
Protein Size (# AA):
997
Molecular Weight:
110kDa
Host:
Rabbit
Clonality:
Polyclonal
Purification:
Affinity Purified
Application:
IHC, WB
Tissue Tool:
Find tissues and cell lines supported by DNA array analysis to express NOS2.
RNA Seq:
Find tissues and cell lines supported by RNA-seq analysis to express NOS2.
Predicted Species Reactivity:
Human
Tested Species Reactivity:
Human
Predicted Homology Based on Immunogen Sequence:
Human: 100%
Complete computational species homology data:
Anti-NOS2 (ARP63000_P050)
Peptide Sequence:
Synthetic peptide located within the following region: PCATSSPVTQDDLQYHNLSKQQNESPQPLVETGKKSPESLVKLDATPLSS
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:
CUL5; SPSB2; NOS2; CALM1; HSP90AA1; CCT7; AGR2; CDIPT; DNAJA2; PSMD14; HUWE1; PSMD6; BAG2; RPL23; AIFM1; DNAJA3; HERC1; SLC7A5; YWHAZ; YWHAG; YWHAE; XRCC5; VCP; UBC; DNAJC7; CCT3; TNFAIP2; TCP1; TCEB2; SSR4; SSBP1; TRIM21; SNRPD3; SNRPB; SMN1; SLC3A2; SKP
Blocking Peptide:
For anti-NOS2 (ARP63000_P050) antibody is Catalog # AAP63000
Datasheets/Manuals:
Printable datasheet for anti-NOS2 (ARP63000_P050) antibody

Product Reviews

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3 Item(s)

02/01/2017 16:24
  • Overall Experience:
  • Quality:
Product Review: NOS2 antibody - N-terminal region (ARP63000_P050) in Human placental tissue using IHC
Product Page for NOS2 antibody - N-terminal region (ARP63000_P050)

Researcher: Dr. Hiten D. Mistry and Anna Czajka, King's College London; Lesia Kurlak, University of Nottingham
Application: IHC
Species + Tissue/Cell type: Human placental tissue
Primary antibody dilution: 1:50
Secondary antibody: Goat anti rabbit-HRP
Secondary antibody dilution: 1:10,000


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
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 tissues 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 50 dilution
Secondary antibody used, dilution, incubation time and temperature: 1 in 10000 goat anti rabbit, 30 minutes at room temperature
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 negative. Positive control: Skeletal muscle.
Please include your detailed tissue preparation and staining procedure/protocol here: Mount paraffin-embedded sections on APES-coated slides. Dry the sections overnight 37 d C.
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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02/01/2017 16:24
  • Overall Experience:
  • Quality:
Product Review: NOS2 antibody-N-terminal region (ARP63000_P050) in Human placental and myometrial tissue lysate using Western Blot
Product Page for NOS2 antibody-N-terminal region (ARP63000_P050)

Researcher: Hiten Mistry, Ania Czajka and Marta Hentschke Ribeiro, King's College London
Application: Western blotting
Species + Tissue/Cell type: Human placental and myometrial tissue lysate
How many ug's of tissue/cell lysate run on the gel:
1: 30 ug human placental tissue lysate
2: 30 ug human placental tissue lysate
3: 30 ug human placental tissue lysate
4: 30 ug human placental tissue lysate
5: 20 ug human myometrial tissue lysate
Primary antibody dilution: 1:500
Secondary antibody: Goat anti-rabbit HRP
Secondary antibody dilution: 1:10000



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. It worked for the tissue lysates and for our positive controls.

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

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?

-20C

Sample Description (please include 1) species type, and 2) cell/tissue type, and 3) how much protein loaded for each lane of your gel):

1. Human, 2. Placental lysate 3. 20-50ug/lane

How many different experimental trials were conducted using the antibody sample?

2-3

How was this sample prepared?

100mg of sample was homogenised in RIPA buffer and treated with protease inhibitors cocktail (SIGMA).

Primary antibody dilution and incubation time:

1:500, overnight incubation

Secondary antibody used and dilution and incubation time:

1:10000, 2-3 hrs

What controls were used in your experiment (positive/negative)?

Positive: myometrial tissue lysate

Please include your detailed WB Procedure/Protocol here:

Equipment: XCell SureLock Mini-cell system with blotting module, gel loading, pipette, tips, mechanical rocker.

1. Loading buffer: Laemlli buffer (Protein loaded (u/5) required to run for each sample)

2. Running buffer (MES SDS RB 20X)

3.Marker (10ug of Novex 4-12% Bis-Tris Gel 1X Mes Running buffer)

4. Gel: 4-12% Bis-Tris gel

5. Electrophorosis Conditions: 1h-1h30min, 175V

6. Transfer Membrane used: Immobilion-P (Millipore) (PVDF)

7. Block in methanol X2 (30 sec/15 sec)

8. Blotting conditions: 1h 30 min, 30V

9. Primary antibody (1:500), overnight Incubation

10. Wash 3 times for 20 minutes at maximum agitation in TBS-Tween after antibodies incubation

11. ECL (3mL per membrane, equal quantities of reagents A and B) for 1 minute

12. Blot analysis: Chemiluminescense (ECL TM)

13. Visualization useing GelLogic 2200 Pro, Carestream Program, from 10sec -30 mindepending on antibody

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02/01/2017 16:24
  • Overall Experience:
  • Quality:
Product Review: NOS2 antibody-N-terminal region (ARP63000_P050) in Human placental and myometrial tissue lysate Western Blot
Product Page for NOS2 antibody-N-terminal region (ARP63000_P050)

Researcher: Hiten Mistry, Ania Czajka and Marta Hentschke Ribeiro, King's College London
Application: Western blotting
Species + Tissue/Cell type: Human placental and myometrial tissue lysate
How many ug's of tissue/cell lysate run on the gel:
1: 50 ug human placental tissue lysate
2: 40 ug human placental tissue lysate
3: 30 ug human placental tissue lysate
4: 20 ug human placental tissue lysate
5: 10 ug human placental tissue lysate
6: 20 ug human myometrial tissue lysate
Primary antibody dilution: 1:500
Secondary antibody: Goat anti-rabbit HRP
Secondary antibody dilution: 1:10000



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. It worked for the tissue lysates and for our positive controls.

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

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?

-20C

Sample Description (please include 1) species type, and 2) cell/tissue type, and 3) how much protein loaded for each lane of your gel):

1. Human, 2. Placental lysate 3. 20-50ug/lane

How many different experimental trials were conducted using the antibody sample?

2-3

How was this sample prepared?

100mg of sample was homogenised in RIPA buffer and treated with protease inhibitors cocktail (SIGMA).

Primary antibody dilution and incubation time:

1:500, overnight incubation

Secondary antibody used and dilution and incubation time:

1:10000, 2-3 hrs

What controls were used in your experiment (positive/negative)?

Positive: myometrial tissue lysate

Please include your detailed WB Procedure/Protocol here:

Equipment: XCell SureLock Mini-cell system with blotting module, gel loading, pipette, tips, mechanical rocker.

1. Loading buffer: Laemlli buffer (Protein loaded (u/5) required to run for each sample)

2. Running buffer (MES SDS RB 20X)

3.Marker (10ug of Novex 4-12% Bis-Tris Gel 1X Mes Running buffer)

4. Gel: 4-12% Bis-Tris gel

5. Electrophorosis Conditions: 1h-1h30min, 175V

6. Transfer Membrane used: Immobilion-P (Millipore) (PVDF)

7. Block in methanol X2 (30 sec/15 sec)

8. Blotting conditions: 1h 30 min, 30V

9. Primary antibody (1:500), overnight Incubation

10. Wash 3 times for 20 minutes at maximum agitation in TBS-Tween after antibodies incubation

11. ECL (3mL per membrane, equal quantities of reagents A and B) for 1 minute

12. Blot analysis: Chemiluminescense (ECL TM)

13. Visualization useing GelLogic 2200 Pro, Carestream Program, from 10sec -30 mindepending on antibody

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