List of IHC/WB antibodies, RT-PCR probes and software used

carmela.giachino

Jun 13, 2023

List of IHC/WB antibodies, RT-PCR probes and software used

DOI

dx.doi.org/10.17504/protocols.io.4r3l27ryjg1y/v1

carmela giachino¹

¹Institut Imagine, INSERM UMR1163 Paris Cite' University, 24 boulevard du Montparnasse, 75015, Paris, France.

carmela giachino

institut imagine

DOI: dx.doi.org/10.17504/protocols.io.4r3l27ryjg1y/v1

Protocol Citation: carmela giachino 2023. List of IHC/WB antibodies, RT-PCR probes and software used. protocols.io https://dx.doi.org/10.17504/protocols.io.4r3l27ryjg1y/v1

License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited

Protocol status: Working

We use this protocol and it's working

Created: March 31, 2023

Last Modified: June 13, 2023

Protocol Integer ID: 79819

Keywords: List of antibodies used for immunohistochemistry, western blot

Funders Acknowledgement:

ASAP-Aligning Science Across Parkinson’s

Grant ID: ASAP-000420

Abstract

This protocol details list of antibodies used for immunohistochemistry and western blot.

Attachments

k4dtb48if.docx

19KB

Materials

Table 1. List of antibodies used for immunohistochemistry and western blot.
 
 
ABCD
ABCD
Antibody nameCompanyDilutionRRID
CCR2, goat polyclonalThermo Fisher Scientific0.388889AB_557978
CD11b, rabbitAbcam0.736111AB_2650514
CD11b, ratBiolegend0.388889AB_312784
CD3, mouseSanta Cruz Biotechnology0.388889AB_2228831
CD4, ratBio-Rad0.25AB_1898234
CD68, ratBio-Rad0.25AB_2074849
DAT, rabbitProteintech0.180556AB_2879116
GFAP, mouseSigma0.215278AB_2827276
GFAP, rabbitsigma0.736111AB_2905668
GSK-3β, mouseECM Biosciences0.180556AB_2115216
HuD+HuC, rabbitAbcam0.319444AB_2864321
Iba-1, goat polyclonalAbcam0.180556AB_870576
Iba-1, rabbit polyclonalWako0.388889AB_2889406
iNOS, mouseCell signaling technology0.180556AB_1078202
LRRK2, rabbitclone 41-2, Abcam by MJFF0.736111AB_2713963
MAP2, rabbitCell Signaling0.388889AB_10693782
NeuN, rabbitAbcam0.736111AB_2716282
Nurr1, goat polyclonalR&D0.25AB_2153894
phospho-alpha-Synuclein
  (Ser129), rabbit monoclonal (clone LS4-1B1)Millipore0.736111AB_673008
phospho-GSK-3β (p-Tyr216), rabbitAbcam0.388889AB_2533691
Phospho-LRRK2-S935, rabbitAbcam by MJFF0.736111AB_2864018
phospho-Tau (pSer396), rabbit polyclonalSigma aldrich0.180556AB_261757
phospho-Tau  (pThr181)Thermo Fisher scientific0.180556AB_1087704
Tau, mouseRayBiotech0.180556AB_11217610
TH, mouseSTEMCELL Technologies0.180556AB_215512
TH, rabbitPel-Freez0.180556AB_2313713
TH, sheepPel-Freez Biologicals0.180556AB_2935637
TH, sheepPel-Freez Biologicals0.736111AB_461070
TUBB3, mouseBiolegend0.319444AB_2313773
α-syn (C-20)-R,
  rabbitSanta Cruz Biotechnology0.180556AB_2192953
α-syn aggregate,
  MJFR-14-6-4-2, rabbitAbcamJan-00AB_2714215
β-actin, rabbitCell Signaling0.736111AB_10694076
 
 
Table 2. List of probes and IDs used for quantitative Real time PCR.

 
 
 
AB
PROBESIDs
Casp6Mm00438053-m1
CCl3Mm 00441258-m1
CxCl10Mm 00445235-m1
CxCl11Mm00444662-m1
Drd1aMm01353211-m1
Drd2Mm00438545-m1
Foxa2Mm01976556-s1
GAPDHMm99999915-g1
GfapMm 00546086-m1
Gp91phoxMm01287743-m1
GSK3bMm00444911-m1
IL-10Mm12088386-m1
IL-17Mm00439618-m1
IL-18Mm00434226-m1
Il1bMm00434228-m1
IL-4Mm00445259-m1
Itgam (Mac1)Mm 00434455-m1
LRRK2Mm00481934_m1
Map2Mm00485231-m1
Nfkb1Mm00476361-m1
Nos 2Mm 00440485-m1
Nurr 1Mm 00443056-m1
Slc6a3 (Dat)Mm 00438388-m1
Tau (Mapt)Mm00521988-m1
ThMm 00447546-m1
TnfMm00443258-m1
Tnfrsf1bMm00441875-m1
α-syn (SNCA)Mm00458965-m1
 

 
 

List of antibodies used for immunohistochemistry and western blot

Table 1. List of antibodies used for immunohistochemistry and western blot.

 
ABCD
Antibody nameCompanyDilutionRRID
CCR2, goat polyclonalThermo Fisher Scientific1:500AB_557978
CD11b, rabbitAbcam1:1000AB_2650514
CD11b, ratBiolegend1:500AB_312784
CD3, mouseSanta Cruz Biotechnology1:500AB_2228831
CD4, ratBio-Rad1:300AB_1898234
CD68, ratBio-Rad1:300AB_2074849
DAT, rabbitProteintech1:200AB_2879116
GFAP, mouseSigma1:250AB_2827276
GFAP, rabbitsigma1:1000AB_2905668
GSK-3β, mouseECM Biosciences1:200AB_2115216
HuD+HuC, rabbitAbcam1:400AB_2864321
Iba-1, goat polyclonalAbcam1: 200AB_870576
Iba-1, rabbit polyclonalWako1:500AB_2889406
iNOS, mouseCell signaling technology1: 200AB_1078202
LRRK2, rabbitclone 41-2, Abcam by MJFF1:1000AB_2713963
MAP2, rabbitCell Signaling1:500AB_10693782
NeuN, rabbitAbcam1:1000AB_2716282
Nurr1, goat polyclonalR&D1:300AB_2153894
phospho-alpha-Synuclein (Ser129), rabbit monoclonal (clone LS4-1B1)Millipore1:1000AB_673008
phospho-GSK-3β (p-Tyr216), rabbitAbcam1:500AB_2533691
Phospho-LRRK2-S935, rabbitAbcam by MJFF1:1000AB_2864018
phospho-Tau (pSer396), rabbit polyclonalSigma aldrich1:200AB_261757
phospho-Tau  (pThr181)Thermo Fisher scientific1:200AB_1087704
Tau, mouseRayBiotech1:200AB_11217610
TH, mouseSTEMCELL Technologies1:200AB_215512
TH, rabbitPel-Freez1:200AB_2313713
TH, sheepPel-Freez Biologicals1:200AB_2935637
TH, sheepPel-Freez Biologicals1:1000AB_461070
TUBB3, mouseBiolegend1:1000AB_2313773
α-syn (C-20)-R, rabbitSanta Cruz Biotechnology1:200AB_2192953
α-syn aggregate, MJFR-14-6-4-2, rabbitAbcam1/5000AB_2714215
β-actin, rabbitCell Signaling1:1000AB_10694076
 

 

 Table 2. List of probes and IDs used for quantitative Real time PCR.
 
 
AB
PROBESIDs
Casp6Mm00438053-m1
CCl3Mm 00441258-m1
CxCl10Mm 00445235-m1
CxCl11Mm00444662-m1
Drd1aMm01353211-m1
Drd2Mm00438545-m1
Foxa2Mm01976556-s1 
GAPDHMm99999915-g1
GfapMm 00546086-m1
Gp91phoxMm01287743-m1
GSK3bMm00444911-m1
IL-10Mm12088386-m1
IL-17Mm00439618-m1
IL-18Mm00434226-m1
Il1bMm00434228-m1
IL-4Mm00445259-m1
Itgam (Mac1)Mm 00434455-m1
LRRK2Mm00481934_m1
Map2Mm00485231-m1
Nfkb1Mm00476361-m1
Nos 2Mm 00440485-m1
Nurr 1Mm 00443056-m1
Slc6a3 (Dat)Mm 00438388-m1
Tau (Mapt)Mm00521988-m1
ThMm 00447546-m1
TnfMm00443258-m1
Tnfrsf1bMm00441875-m1
α-syn (SNCA)Mm00458965-m1
 
 

 Table 3 List of software 
ABC
  Software name    Version    URL  
  ImageJ    1.53t    https://imagej.net/  
  Luminex xPONENT    4.3       https://www.luminexcorp.com/xponent/  
  Bio-Plex Manager    6.2       http://www.bio-rad.com/en-us/product/bio-plex-manager-software-standard-edition     
  ImageQuantity One    4.6    http://www.bio-rad.com/en-us/product/quantity-one-1-d-analysis-software  
  GraphPad Prism    9    http://www.graphpad.com/  
 

A	B	C	D
A	B	C	D
Antibody name	Company	Dilution	RRID

CCR2, goat polyclonal	Thermo Fisher Scientific	0.388889	AB_557978
CD11b, rabbit	Abcam	0.736111	AB_2650514
CD11b, rat	Biolegend	0.388889	AB_312784
CD3, mouse	Santa Cruz Biotechnology	0.388889	AB_2228831
CD4, rat	Bio-Rad	0.25	AB_1898234
CD68, rat	Bio-Rad	0.25	AB_2074849
DAT, rabbit	Proteintech	0.180556	AB_2879116
GFAP, mouse	Sigma	0.215278	AB_2827276
GFAP, rabbit	sigma	0.736111	AB_2905668
GSK-3β, mouse	ECM Biosciences	0.180556	AB_2115216
HuD+HuC, rabbit	Abcam	0.319444	AB_2864321
Iba-1, goat polyclonal	Abcam	0.180556	AB_870576
Iba-1, rabbit polyclonal	Wako	0.388889	AB_2889406
iNOS, mouse	Cell signaling technology	0.180556	AB_1078202
LRRK2, rabbit	clone 41-2, Abcam by MJFF	0.736111	AB_2713963
MAP2, rabbit	Cell Signaling	0.388889	AB_10693782
NeuN, rabbit	Abcam	0.736111	AB_2716282
Nurr1, goat polyclonal	R&D	0.25	AB_2153894
phospho-alpha-Synuclein (Ser129), rabbit monoclonal (clone LS4-1B1)	Millipore	0.736111	AB_673008
phospho-GSK-3β (p-Tyr216), rabbit	Abcam	0.388889	AB_2533691
Phospho-LRRK2-S935, rabbit	Abcam by MJFF	0.736111	AB_2864018
phospho-Tau (pSer396), rabbit polyclonal	Sigma aldrich	0.180556	AB_261757
phospho-Tau (pThr181)	Thermo Fisher scientific	0.180556	AB_1087704
Tau, mouse	RayBiotech	0.180556	AB_11217610
TH, mouse	STEMCELL Technologies	0.180556	AB_215512
TH, rabbit	Pel-Freez	0.180556	AB_2313713
TH, sheep	Pel-Freez Biologicals	0.180556	AB_2935637
TH, sheep	Pel-Freez Biologicals	0.736111	AB_461070
TUBB3, mouse	Biolegend	0.319444	AB_2313773
α-syn (C-20)-R, rabbit	Santa Cruz Biotechnology	0.180556	AB_2192953
α-syn aggregate, MJFR-14-6-4-2, rabbit	Abcam	Jan-00	AB_2714215
β-actin, rabbit	Cell Signaling	0.736111	AB_10694076


	A	B
	PROBES	IDs
	Casp6	Mm00438053-m1
	CCl3	Mm 00441258-m1
	CxCl10	Mm 00445235-m1
	CxCl11	Mm00444662-m1
	Drd1a	Mm01353211-m1
	Drd2	Mm00438545-m1
	Foxa2	Mm01976556-s1
	GAPDH	Mm99999915-g1
	Gfap	Mm 00546086-m1
	Gp91phox	Mm01287743-m1
	GSK3b	Mm00444911-m1
	IL-10	Mm12088386-m1
	IL-17	Mm00439618-m1
	IL-18	Mm00434226-m1
	Il1b	Mm00434228-m1
	IL-4	Mm00445259-m1
	Itgam (Mac1)	Mm 00434455-m1
	LRRK2	Mm00481934_m1
	Map2	Mm00485231-m1
	Nfkb1	Mm00476361-m1
	Nos 2	Mm 00440485-m1
	Nurr 1	Mm 00443056-m1
	Slc6a3 (Dat)	Mm 00438388-m1
	Tau (Mapt)	Mm00521988-m1
	Th	Mm 00447546-m1
	Tnf	Mm00443258-m1
	Tnfrsf1b	Mm00441875-m1
	α-syn (SNCA)	Mm00458965-m1

A	B	C
Software name	Version	URL
ImageJ	1.53t	https://imagej.net/
Luminex xPONENT	4.3	https://www.luminexcorp.com/xponent/
Bio-Plex Manager	6.2	http://www.bio-rad.com/en-us/product/bio-plex-manager-software-standard-edition
ImageQuantity One	4.6	http://www.bio-rad.com/en-us/product/quantity-one-1-d-analysis-software
GraphPad Prism	9	http://www.graphpad.com/

Public workspaceList of IHC/WB antibodies, RT-PCR probes and software used

List of IHC/WB antibodies, RT-PCR probes and software used