CDC42EP1 Antibody

Images

 
Genetic Strategies: Western Blot: CDC42EP1 Antibody [NBP1-88379] - HeLa whole cell lysate. Lanes 1 and 4: HeLa cells transfected with control non-targeting siRNA. Lanes 2 and 3: HeLa cells transfected with two ...read more
Immunocytochemistry/ Immunofluorescence: CDC42EP1 Antibody [NBP1-88379] - Confocal Imunofluorescent images of HeLa cells fixed with PFA and stained with Cdc42EP1 antibody at 1:200 (magenta), and phalloidin-Alexa 488 ...read more
Immunohistochemistry-Paraffin: CDC42EP1 Antibody [NBP1-88379] - Staining of human stomach shows strong cytoplasmic positivity in glandular cells.
Immunocytochemistry/ Immunofluorescence: CDC42EP1 Antibody [NBP1-88379] - Staining of human cell line A-431 shows localization to plasma membrane & actin filaments. Antibody staining is shown in green.
Immunoprecipitation: CDC42EP1 Antibody [NBP1-88379] - Lanes 1 and 5: Marker. Lanes 2-4 HeLa cell input controls. Lane 6: IP using 2 mg of Control Rabbit IgG cell lysate. Lane 8: IP using 2 mg Cdc42EP1 cell lysate and ...read more

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB, ICC/IF, IHC, IHC-P, IP, KD
Clonality
Polyclonal
Host
Rabbit
Conjugate
Unconjugated
Validated by:
     

Genetic Strategies

   

Order Details

CDC42EP1 Antibody Summary

Immunogen
This antibody was developed against Recombinant Protein corresponding to amino acids: SGFCTISRLPRSEKPHDRDRDGSFPSEPGLRRSDSLLSFRLDLDLGPSLLSELLGVMSLPEAPAAETPAPAANPPAPTANPTGPAANPPATTANPPAPAANPSAPAATPTGPAANPPAPAASSTPHGHCPNGV
Isotype
IgG
Clonality
Polyclonal
Host
Rabbit
Gene
CDC42EP1
Purity
Immunogen affinity purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Immunocytochemistry/Immunofluorescence 0.25-2 ug/ml
  • Immunohistochemistry 1:50 - 1:200
  • Immunohistochemistry-Paraffin 1:50 - 1:200
  • Immunoprecipitation
  • Knockdown Validated
  • Western Blot 0.04-0.4 ug/ml
Application Notes
For IHC-Paraffin, HIER pH 6 antigen retrieval is recommended. ICC/IF fixation Permeabilization, Use PFA/Triton X-100. CDC42EP1 antibody validated for ICC/IF, IP,WB from verified customer reviews.
Control Peptide
CDC42EP1 Protein (NBP1-88379PEP)
Reviewed Applications
Read 3 Reviews rated 5
using
NBP1-88379 in the following applications:

Packaging, Storage & Formulations

Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Buffer
PBS (pH 7.2) and 40% Glycerol
Preservative
0.02% Sodium Azide
Purity
Immunogen affinity purified

Alternate Names for CDC42EP1 Antibody

  • Binder of Rho GTPases 5
  • Borg5
  • BORG5cdc42 effector protein 1
  • CDC42 effector protein (Rho GTPase binding) 1
  • CEP155 kDa bone marrow stromal/endothelial cell protein
  • MSE55MGC15316
  • serum constituent protein
  • Serum protein MSE55

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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Publications for CDC42EP1 Antibody (NBP1-88379) (0)

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Reviews for CDC42EP1 Antibody (NBP1-88379) (3) 53

Average Rating: 5
(Based on 3 reviews)
We have 3 reviews tested in 1 species: Human.

Reviews using NBP1-88379:
Filter by Applications
ICC
(1)
IP
(1)
WB
(1)
All Applications
Filter by Species
Human
(3)
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Images Ratings Applications Species Date Details
Immunocytochemistry CDC42EP1 NBP1-88379
Enlarge
5
reviewed by:
Stephen Terry
ICC Human 10/25/2018
View

Summary

ApplicationImmunocytochemistry
Sample Tested hela cell
SpeciesHuman
LotA96959

Comments

CommentsCells fixed with 4% PFA for 20 minutes room temperature.

Fixed cells incubated with with 0.5% triton x100 in 1xPBS for 5 minutes room.

Washed twice in blocking buffer (1% BSA 1xPBS, 20mM glycine).

Washed once with blocking buffer left for 1hour room temperature.

incubated with Anti Cdc42EP1 antibody in blocking buffer 1:200 for 20 hours at 4 degrees.

Washed 3 x 5 minutes in blocking buffer

Incubated with Anti rabbit-Alexa 594 conjugated antibody (Jackson immunoscience) for 2 hours room temperature.

Washed 3 x 5 minutes in blocking buffer

Washed 1 x 5 minutes in PBS

Imaged with 60X oil objective on Nikon Eclipse Ti Inverted Fast confocal - Equipped with a Yokogawa CSU-X1 spinning disk unit and okogawa CSU-1 disk head and Andor Neo sCMOS camera.
Immunoprecipitation CDC42EP1 NBP1-88379
Enlarge
5
reviewed by:
Stephen Terry
IP Human 08/20/2018
View

Summary

ApplicationImmunoprecipitation
Sample Tested hela cell
SpeciesHuman
LotA96959

Comments

CommentsVortex tube of mag protein G beads (NEB)

Take 30 μl put in tube

Add 1 ml of 1XPBS to wash

Put on mag rack discard PBS

Suspend beads in 60 μl of 1XPBS

Add between 2mg of primary IP antibody to beads

Put on rotating wheel at 4°C for 20h

Lysis of cells

Put all microfuge tubes that you are going to use on ice

Cool centrifuge to 4°C

Take HeLa cells grown on 10cm dishes to about 70% confluence

Aspirate media wash twice with 10ml cold 1xPBS

Add 1 ml of lysis buffer + 1xphosphatase + 1xprotease inhibtors (Roche) to each 10cm plate

Scrape cells off plate

Syringe 7-8 times with 25 G needle

Transfer to microfuge tube on ice

pre clear lysate by adding 60 μl of mag protein G beads (NEB) to each 1 ml of lysate

Leave on ice for 10 min inverting several times.

Spin tubes in cool centrifuge max speed 13K rpm for 5 min

Put tubes in mag rack pipette off supernatant

Save 50 μl of one of the samples to use as an input lane

add 50 μl of 3X sample buffer +0.15M DTT to this boil for 5 min store -20°C

save as input -load 20ul of this on gel

Add 1ml of pre cleared lysate to each of the protein G –Antibody conjugates prepared day before

Put on rotating wheel in cold room for 2h

Put tubes on mag rack

Wash once with 1ml cold 0.5% triton x100 PBS

Put beads on rack remove liquid

Wash 3X more with 1ml cold 1XPBS

Suspend beads in 80 μl of 3X of 3X sample buffer +0.3MDTT boil 10 min

Put on mag rack

Transfer liquid to new tube

Load 15 μl on gel along with 20 μl of inputs


Gel run out on 10% SDS PAGE gel in MOPS buffer
Transfered 100V 2hours 4 degrees nitrocellulose
membrane blocked 5% milk PBST 1 hour
incubated overnight with Cdc42EP1 antibody NBP1-88379 1:2000 in 5% milk PBST
Washed 3X PBST
Incubated with secondary anti rabbit HRP con light chain specific antibody 1:5000
(Jackson immunoresearch 211-032-171) in for 1hour 5% milk PBST
Washed 3X PBST
washed once 1X PBS
ECL prime kit GE healthcare used for detection of chemilumincence
Western Blot CDC42EP1 NBP1-88379
Enlarge
5
reviewed by:
Stephen Terry
WB Human 08/03/2018
View

Summary

ApplicationWestern Blot
Sample Tested Hela whole cell lysate
SpeciesHuman
LotA96959

Comments

CommentsFor siRNA blot two different siRNAs were transfected at 40nM, targeting cdc42ep1 or non targeting control siRNA cells were lysed after 72 hours post tranfection
Protein lysates were lysed in 1.5X Lammeli sample buffer boiled at 100 degrees for 5 minutes
10ug of protein loaded on 10% SDS PAGE gel run in MOPS buffer
I used the antibody at 1:2000 in 5% milk TBST incubation overnight 4 degrees
antibody gives a band at -55kD that reduces in RNAi
Membranes were reprobed with GAPDH antibody as a loading control

Product General Protocols

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Video Protocols

WB Video Protocol
ICC/IF Video Protocol

FAQs for CDC42EP1 Antibody (NBP1-88379) (0)

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Secondary Antibodies

 

Isotype Controls

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Recent Reviews

5
5
3
4
0
3
0
2
0
1
0

Stephen Terry
10/25/2018
Application: ICC
Species: Human

Stephen Terry
08/20/2018
Application: IP
Species: Human

Stephen Terry
08/03/2018
Application: WB
Species: Human

Bioinformatics

Gene Symbol CDC42EP1