antibodies that work in tribolium castaneum
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Antibodies that Work In Tribolium Castaneum. Kevin Burns 5/20/2011. Notes on this Document. For antibodies I’ve used personally, the titles contain the following information Original antigen used to produce antibody A rough guess at the stage/stages of Tc shown

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notes on this document
Notes on this Document
  • For antibodies I’ve used personally, the titles contain the following information
    • Original antigen used to produce antibody
    • A rough guess at the stage/stages of Tc shown
    • Antibody procurement info, e.g. DSHB, Supplier, or Lab
    • Host species and working dilution
  • I have often thrown in co-stains to further illustrate morphology and to make things look cool. However, mostly this is just confusing.
    • KT817 is an enhancer trap line from Sue Brown that marks oenocytes. It is also cool for marking A1-A8
  • The second half of this document in-exhaustively covers antibodies that have been used in published works, normally on beetle, but occasionally on other arthropods, please update me if you find any new ones (Kevin.Burns@cchmc.org)
  • There is a PDF and a Powerpoint version of this document (The hyperlinks work in the PPT), if your email server does not support attachments of such & such size, I will post this document on http://burnska08.wordpress.com/kevin-burns-home-page/
index
Index
  • AbdA (+UBX)
  • AntP
  • Futsch (22c10)
  • HNF4
  • Notch Intracellular Domain
  • Orthodenticle (GP / Rat)
  • Phospho-Histone3
  • Phospho-MAPK
  • Spalt
  • Trachea (a2A12)
  • UBX (see AbdA)

Index of Other’s Results

dm abd a ubx pre hatching 64h at 30c dshb mouse monoclonal 1 5
Dm Abd-A/UBXPre-Hatching, ~64h at 30CDSHB: Mouse Monoclonal, 1:5

UBX/AbdA

DAPI

KT817-YFP

UBX/AbdA

Index for KB’s Antibodies

dm antp germband retraction 30h @30c dshb mouse monoclonal 1 25
Dm AntPGermband retraction 30h @30CDSHB: Mouse Monoclonal, 1:25+

KT817-YFP

DAPI

AntP

AntP

NICD

Please advise naïve young investigators that AntP is not just expressed in thoracic segments but also in the CNS

Index for KB’s Antibodies

dm en inv dorsal closure leg articulation 36h dshb 4d9 mouse monoclonal 1 25
Dm En/InvDorsal closure/Leg articulation (36h)DSHB: 4D9Mouse Monoclonal, 1:25

En/Inv

Spalt

En/Inv

Index for KB’s Antibodies

tc engrailed cocalico gebelein coming in july rat guinea pig
Tc EngrailedCocalico/Gebelein, coming in JulyRat/Guinea Pig

Kevin Burns is making this in Brian Gebelein’s lab in order to do some co-stains with the multitude of mouse antibodies

Index for KB’s Antibodies

dm futsch dorsal closure 48h ventral view dshb 22c10 mouse monoclonal 1 50
Dm FutschDorsal closure/48h? (Ventral View)DSHB: 22c10Mouse Monoclonal, 1:50

22c10

Index for KB’s Antibodies

hnf4 48h @ 30c cocalico gebelein rat serum 1 1000
HNF448h @ 30CCocalico/Gebelein: Rat Serum, 1:1000

KT817-YFP

HNF4

p

HNF4

Mesodermal staining (arrowheads), continues underneath more superficial oenocyte staining (e.g. red circle)

p = pleuropod in A1

Index for KB’s Antibodies

notch intracellular domain germband extension dshb c17 9c6 mouse monoclonal 1 5
Notch Intracellular DomainGermband extensionDSHB: C17.9C6Mouse Monoclonal 1:5

NICD

NICD

DAPI

Index for KB’s Antibodies

dm otd full length mix sample cocalico cook rat serum 1 500
Dm OTD full lengthMix sampleCocalico/Cook: Rat Serum, 1:500

OTD

Index for KB’s Antibodies

I am conducting another stain to see if this detects the OTD that replaces bicoid in early patterning…

phospho histone 3 full extension millipore 05 806 mouse 1 1000
phospho-Histone 3Full extensionMillipore: 05-806Mouse, 1:1000

pH3

pH3

DAPI

Index for KB’s Antibodies

di phosphorylated mapk post gastrulation and full germband sigma m 8159 mouse monoclonal 1 50
Di-Phosphorylated MAPKpost-gastrulation and full germbandSigma: M-8159Mouse Monoclonal, 1:50

pERK

pERK

Index for KB’s Antibodies

dm spalt major 24h 36h @ 30c cocalico gebelein rabbit serum 1 1500
Dm Spalt Major24h, 36h @ 30CCocalico/Gebelein: Rabbit Serum, 1:1500

Spalt

Spalt

pERK

Spalt

Spalt

KT-817-YFP

Index for KB’s Antibodies

dm unknown tracheal system pre hatching dshb a2a12 mouse igm monoclonal 1 25
Dm unknown (tracheal System)Pre-HatchingDSHB: a2A12 Mouse IGM Monoclonal, 1:25

This stain was achieved with IgG secondary antibodies, my guess is secondaries designed against IgM would fare much better

Index for KB’s Antibodies

NICD

a2A12

DAPI

table of kevin s antibodies
Table of Kevin’s Antibodies

Index for KB’s Antibodies

index of other people s
Index of Other People’s
  • Caudal
  • Caspase3
  • Cx (scr)
  • DC0(mushroom body etc)
  • DLL
  • engrailed 4F11 from DSHB
  • Even-skipped
  • Nubbin (Drosophila + Crustacean + Chelicerates)
  • Prd, Gooseberry, Gooseberry-Neuro (Pairberry)
  • pSMAD
  • pY
  • Synapsin 1

Index for KB’s Antibodies

caudal shulz tautz 1998 dev genes evolution
CaudalShulz&Tautz 1998 Dev Genes & Evolution

Generation of an antibody

Part of the cadA ORF (open reading frame, corresponding to amino

acids 79–249) was amplified by PCR and cloned in frame into

the EcoRV site of the pRSET-C (Invitrogen) vector. The

Index of Other’s Results

caspase 3 aranda tautz2009 dev genes evol
Caspase 3Aranda&Tautz2009 Dev Genes Evol

For apoptosis detection using the anti cleaved caspase3 antibody (Cell Signalling Technology, Inc.)

Index of Other’s Results

cx scr shippy denell 2006 genetics
Cx (Scr)Shippy & Denell 2006 Genetics

A cross-reacting polyclonal antibody to Drosophila Scr, α-DmScr (a gift from Thomas Kaufman), was used to detect Cx

Index of Other’s Results

dc0 zhao farris 2008
DC0Zhao&Farris 2008

Anti-DC0 primary antibody, a polyclonal antibody

against the catalytic subunit of Drosophila melanogaster

protein kinase A, was a gift provided by Dr. Daniel Kalderon.

This antibody has been shown to have a high affinity

for mushroom body intrinsic neurons in a range of insect

species (Skoulakis et al., 1993; Farris, 2005a,b).

Index of Other’s Results

dll e g panganiban carroll 1997 pnas
DLLe.g. Panganiban&Carroll 1997 PNAs
  • Butterfly Staining…

Production and characterization of the Dll homeodomain antibody was described in Panganiban et al. (4)

Index of Other’s Results

engrailed 4f11 alternate dshb clone e g pueyo couso2008 pnas
Engrailed 4F11 (alternate DSHB Clone)e.g. Pueyo&Couso2008 PNAS

This is cochroach, but I swear I’ve seen beetle people use 4f11 instead of 4D9

Index of Other’s Results

even skipped brown denell 1997
Even SkippedBrown & Denell 1997

Choe & Brown 2006

Grasshopper antigen, see also Patel 1994

Index of Other’s Results

nubbin li popadic 2004 evolution development
NubbinLi & Popadic 2004 Evolution& Development

A monoclonal antibody that recognizes a conserved epitope in the Nub/pdm protein has been generated and used successfully in a number of crustacean and chelicerate species (Averof and Cohen 1997; Abzhanov and Kaufman 2000; Damen et al. 2002). Unfortunately, this antibody does not recognize insect nub orthologs except for that of Drosophila.

Index of Other’s Results

phospho tyrosine nunes da fonseca roth 2008 dev cell
Phospho-TyrosineNunes da fonseca& Roth 2008 Dev Cell

Anti-phosphotyrosine antibody staining marks the cell outlines (red), DAPI (blue)

Index of Other’s Results

synapsin 1 dreyer schactner2010
Synapsin1Dreyer&Schactner2010

The monoclonal primary antibody from mouse against a fusion protein consisting of a glutathione-S-transferase and the first amino acids of the presynaptic vesicle protein synapsin I coded by its 5′-end (SYNORF1, Klagges et al., 1996 )

Index of Other’s Results

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