- Tested Species Reactivity:
- Predicted Species Reactivity:
- Cow, Human, Mouse, Pig, Rat
- Product Format:
- Liquid. Purified antibody supplied in 1x PBS buffer with 0.09% (w/v) sodium azide and 2% sucrose.
- IHC, WB
- 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.
- Gene Symbol:
- Official Gene Full Name:
- Caveolin 3
- NCBI Gene Id:
- Protein Name:
- Swissprot Id:
- Protein Accession #:
- Nucleotide Accession #:
- Alias Symbols:
- LQT9, VIP21, LGMD1C, VIP-21
- Replacement Item:
- This antibody may replace item sc-30752 from Santa Cruz Biotechnology.
- Description of Target:
- CAV3 is a caveolin family member, which functions as a component of the caveolae plasma membranes found in most cell types. Caveolin proteins are proposed to be scaffolding proteins for organizing and concentrating certain caveolin-interacting molecules. Mutations identified in this gene lead to interference with protein oligomerization or intra-cellular routing, disrupting caveolae formation and resulting in Limb-Girdle muscular dystrophy type-1C (LGMD-1C), hyperCKemia or rippling muscle disease (RMD). Alternative splicing has been identified for this locus, with inclusion or exclusion of a differentially spliced intron. In addition, transcripts utilize multiple polyA sites and contain two potential translation initiation sites.
- Protein Size (# AA):
- Molecular Weight:
- Affinity Purified
- Tissue Tool:
- Find tissues and cell lines supported by DNA array analysis to express CAV3.
- RNA Seq:
- Find tissues and cell lines supported by RNA-seq analysis to express CAV3.
- The immunogen is a synthetic peptide directed towards the N terminal region of human CAV3
- Predicted Homology Based on Immunogen Sequence:
- Cow: 100%; Human: 100%; Mouse: 84%; Pig: 92%; Rat: 84%
- Complete computational species homology data:
- Anti-CAV3 (AVARP09021_P050)
- Peptide Sequence:
- Synthetic peptide located within the following region: MMAEEHTDLEAQIVKDIHCKEIDLVNRDPKNINEDIVKVDFEDVIAEPVG
- Batch dependent within range: 100 ul at 0.5 - 1 mg/ml
- Protein Interactions:
- KCNMA1; PTGES3; SCN5A; PCBP1; NEDD4L; KCNH2; SUMO2; SUMO3; ADRB2; JPH2; SLC22A11; DYSF; PFKM; SLC8A1; NOS1; INSR; GNAS; EGFR; DAG1; PDGFRB; PDGFRA;
- Blocking Peptide:
- For anti-CAV3 (AVARP09021_P050) antibody is Catalog # AAP30146 (Previous Catalog # AAPP00303)
- Printable datasheet for anti-CAV3 (AVARP09021_P050) antibody
- Target Reference:
- Alias,L., et al., (2004) Neuromuscul. Disord. 14 (5), 321-324
Researcher: Dr. Hiten D. Mistry and Anna Czajka, King's College London; Lesia Kurlak, University of Nottingham
Species + Tissue/Cell type: Human placental tissue
Primary antibody dilution: 1:50
Secondary antibody: Goat anti rabbit-HRP
Secondary antibody dilution: 1:10,000
|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 200 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 dependson 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
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: 20 ug human myometrial tissue lysate
Primary antibody dilution: 1:500
Secondary antibody: Goat anti-rabbit HRP
Secondary antibody dilution: 1:10000
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 did worked for tissue lysates of interest and for our positive control.
Would you use this antibody in future experiments?
Have you used another antibody which has worked in your application?
Do you believe the information about the reagent on Aviva's website is correct?
If the antibody works, do you plan to use it in future experiments or to publish your data? Why or why not?
How did you store the antibody after re-suspension?
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?
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