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exd antibody - C-terminal region (ARP47897_T100)

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100 ul

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Conjugation Options

ARP47897_T100-FITC Conjugated

ARP47897_T100-HRP Conjugated

ARP47897_T100-Biotin Conjugated

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Gene Symbol:
exd
NCBI Gene Id:
32567
Official Gene Full Name:
Extradenticle
Protein Name:
Homeobox protein extradenticle
Swissprot Id:
P40427
Protein Accession #:
NP_727924
Nucleotide Accession #:
NM_167479
Alias Symbols:
Dmel_CG8933, CG8933, DExd, Dm-EXD, Dpbx, EXD, td48, unnamed, anon-EST:fe1H3, Dmel\CG8933, Exd, l(1)IV, Pbx1
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.
Protein Size (# AA):
376
Molecular Weight:
42kDa
Host:
Rabbit
Clonality:
Polyclonal
Purification:
Protein A purified
Application:
WB
Tissue Tool:
Find tissues and cell lines supported by DNA array analysis to express exd.
RNA Seq:
Find tissues and cell lines supported by RNA-seq analysis to express exd.
Immunogen:
The immunogen is a synthetic peptide corresponding to a region of Fruit fly
Species Reactivity:
Cow, Dog, Guinea Pig, Horse, Human, Mouse, Rabbit, Rat, Zebrafish
Predicted Homology Based on Immunogen Sequence:
Cow: 93%; Dog: 93%; Guinea Pig: 93%; Horse: 93%; Human: 100%; Mouse: 93%; Rabbit: 93%; Rat: 100%; Zebrafish: 93%
Complete computational species homology data:
Anti-exd (ARP47897_T100)
Peptide Sequence:
Synthetic peptide located within the following region: EAKEELARKCGITVSQVSNWFGNKRIRYKKNIGKAQEEANLYAAKKAAGA
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:
CG31957; CG18764; Fer2LCH; CG6454; fzo; ATPsyn-d; hth; CG7443; Dfd; Sec8; CG32425; CG32176; tamo; RpS11; en; nito; E2f2; CG13962; CG7231; Rack1; sip2; Taf12L; dod; Bap60; CG2972; CG12661; MED22; ftz;
Blocking Peptide:
For anti-exd (ARP47897_T100) antibody is Catalog # AAP47897 (Previous Catalog # AAPP27341)
Datasheets/Manuals:
Printable datasheet for anti-exd (ARP47897_T100) antibody
 
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02/01/2017 16:23
  • Overall Experience:
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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
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