exd antibody - C-terminal region (ARP47897_T100)

Catalog#: ARP47897_T100

• Size: 100ug

Drosophila

Drosophila

r-Exd

• Description of Target: As a transcription factor, exd acts with the selector homeodomain proteins altering the regulation of downstream target genes such as wingless, teashirt and decapentaplegic. Thus exd affects segmental identity.
• Gene Symbol: exd
• Official Gene Full Name: Extradenticle
• Alias Symbols: Dmel_CG8933; CG8933; DExd; Dm-EXD; Dpbx; EXD; td48; unnamed; anon-EST:fe1H3; Dmel\CG8933; Exd; l(1)IV; Pbx1
• Protein Accession#: NP_727924
• Nucleotide Accession#: NM_167479
• Swissprot Id: P40427
• Host: Rabbit
• Clonality: Polyclonal
• Protein Size (# AA): 376
• Molecular Weight: 42kDa
• Application: WB
• Partner Proteins: YWHAG
• Immunogen: The immunogen for anti-exd antibody: synthetic peptide corresponding to a region of Fruit fly
• Product Format: Lyophilized powder
• Purification: Protein A purified
• Predicted Homology Based on Immunogen Sequence: Starlet sea anemone:100%; African malaria mosquito:100%; Wandering spider:100%; Yellowfever mosquito:100%; Harpegnathos saltator:100%; Camponotus floridanus:100%; Body louse:100%; edible Chinese black ant:100%; Red flour beetle:100%; Southern house mosquito:100%; Trichoplax reptans:100%; Acorn worm:100%; Black-legged tick:100%; Savannah tsetse fly:100%; Bovine:92%; Transparent sea squirt:92%; Green puffer:92%; African clawed frog:92%; Common tree shrew:92%; Malayan flying lemur:92%; Ring-tailed lemur:92%; Philippine tarsier:92%; Southern owl monkey:92%; Crab-eating macaque:92%; Human:100%; Rainbow trout:92%; Tunicate:92%; Dog:92%; Zebrafish:92%; Pig:92%; Filarial nematode worm:92%; Mouse:92%; Rat:92%; Western clawed frog:92%; Chicken:92%; Blood fluke:92%; Florida lancelet:92%; Atlantic salmon:92%; Eye worm:90%; 
• Peptide Sequence: Synthetic peptide located within the following region: EAKEELARKCGITVSQVSNWFGNKRIRYKKNIGKAQEEANLYAAKKAAGA
• Blocking Peptide: For anti-exd antibody is Catalog # AAP47897 (Previous Catalog # AAPP27341)
• Reconstitution and Storage: Add 100 μl of distilled water. Final anti-exd antibody concentration is 1 mg/ml in PBS buffer. For longer periods of storage, store at -20°C. Avoid repeat freeze-thaw cycles.

Product Protocols: exd antibody tested with Human Drosophila (ARP47897_T100)

Aviva Systems Biology is the original manufacturer of this exd antibody (ARP47897_T100)

Click here to view the exd antibody Western Blot Protocol

Product Datasheet Link: exd antibody (ARP47897_T100)

WB Suggested Anti-exd Antibody Titration: 5.0ug/ml
Positive Control: Drosophila

Western Blot image:


Description of Target: As a transcription factor, exd acts with the selector homeodomain proteins altering the regulation of downstream target genes such as wingless, teashirt and decapentaplegic. Thus exd affects segmental identity.

Questions pertaining to this data can be directed to techsupport@avivasysbio.com

Aviva Systems Biology’s exd antibody (ARP47897_T100) has been tested using other cell lysates and tissues. To obtain more data about this antibody please email us at info@avivasysbio.com.

To order by phone call us at (888) 880-0001, fax us at (858) 552-6975 or send an email to info@avivasysbio.com. Aviva manufactures this antibody so we can offer the best price. Please contact us to request pricing information.

All of Aviva’s products are guaranteed for the applications and experimental sample types mentioned in the datasheet below. Are you curious if this product will work for you? Please contact us at techsupport@avivasysbio.com

Product Review: exd antibody-C-terminal region (ARP47897_T100) in His-tag cell free protein using Western Blot

Product Page for exd antibody-C-terminal region (ARP47897_T100)

Researcher: Kimberly Kaufman, Univeristy of Wisconsin-Madison
Application: Western blotting
Species+tissue/cell type: Drosophila
How many ug’s of tissue/cell lysate run on the gel:
1: E. coli purified Exd (no tag)
2: Cell free expressed Exd (His tag)
Primary antibody dilution: 1:1000
Secondary antibody: Anti-rabbit HRP
Secondary antibody dilution: 1:5000

Questionnaire:

How do Aviva’s reagents play a role in your experimental goals?

I was looking to observe Exd by Western blot, so I used this as my primary antibody (secondary =anti-rabbit HRP conjugate).

How would you rate this antibody on a scale from 1-5 (5=best) and why?

5, it performed exactly as expected.

Would you use this antibody in future experiments?

Definitely.

Have you used another antibody which has worked in your application?

I usually use an anti-His antibody from GE Healthcare.

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?

I only used it to further confirm the presence of Exd, which had previously been demonstrated by the anti-His antibody. However, if my project took a different direction, this antibody should be good for ChIP.

Sample Description:

I tested two forms of Exd- both were visible using this antibody.

Species, Tissue/Cell Type:

Drosophila

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

Two, but I messed up the first Western (the one that worked is in the other e-mail attachment).

What type of experimental sample are you using and how did you prepare it?

Two Exd constructs- one full length expressed in a cell-free expression system (not purified), one with the first four helices purified from E. coli.

Primary used and dilution:

Anti-Exd from Aviva (#ARP47897), dissolved to 1 mg/mL used in 1:1000 dilution

Secondary used and dilution:

Goat anti-rabbit-HRP; 1:5000 dilution.

Experimental Procedure/Protocols:

1. Ran a 15% SDS-PAGE to separate proteins

2. Transferred proteins to nitrocellulose membrane (please let me know if you need more details on this step).

3. Blocked membrane with 5% milk/TBStween(=TBSt=0.05% tween in TBS), 1 h at room temp (all at room temp)

4. Rinsed membrane with 10 mL TBSt, immediately discarded

5. Washed three times at 10 mL for 10 min each

6. Primary antibody solution for 1 h

10 uL anti-Exd (1 mg/mL)

0.2 g BSA

200 uL 100 mM NaN3

9.8 mL TBSt

this solution can be reused for a few months when stored at 4oC

7. Repeated steps four and five

8. Secondary antibody solution for 1 h

2 uL Goat anti-Rabbit+HRP conjugate

0.2 g milk

10 mL TBSt

9. Repeated steps four and five

10. Treated with reagents to light up the HRP- exposed to film

What controls were used in your experiment? Please include your positive control:

I used two different Exd samples from two different sources. The E. coli expression was Exd-C (homeodomain with 4th helix) and had no tag. The cell free expression was full length Exd and had a His tag. Both proteins are visible with the anti-Exd antibody, only the cell free protein is visible with my anti-His antibody.

How did you store the antibody after re-suspension?

4oC