First immunohistochemical evidence of human tendon repair following stem cell injection: A case report and review of literature

Table 1 Characteristics of the antibodies used in this study

Aggrecan
Immunoglobulin isotype/clone status	IgG1/mouse, monoclonal
Catalog no./provider	12/21/1-C-6/DSHB1
Demasking of antigen	Protease XIV and Chondroitinase AC
Blocking	Bloxall SP-60002, NHS S-20002, 1:20
Dilution and incubation parameters	1:5, room temperature, 4 h
Secondary antibody used	HaM IgG BA-20002, 1:200
CD34
Immunoglobuline isotype/clone status	IgG1/mouse, monoclonal
Catalog no./provider	QBEnd-10/Thermo Scientific (Waltham, MA, United States)
Demasking of antigen	N/A
Blocking	3% H2O2 in methanol, NGS S-10002 5%
Dilution and incubation parameters	1:900, 4 °C, overnight
Secondary antibody used	GaM IgG BA-92002, 1:200, in 2% BSA/PBS
CD68
Immunoglobuline isotype/clone status	KP1- IgG1 kappa/mouse, monoclonal
Catalog no./provider	M0814/Dako (Glostrup, Denmark)
Demasking of antigen	Boiling in citrate buffer (pH 6)
Blocking	Bloxall SP-60002, NHS S-20002, 1:20
Dilution and incubation parameters	1:100, 4 °C, overnight
Secondary antibody used	HaM IgG BA-20002, 1:200
Ki-67
Immunoglobuline isotype/clone status	MIB-1 IgG1 kappa/mouse, monoclonal
Catalog no./provider	M7240/Dako (Glostrup, Denmark)
Demasking of antigen	Boiling in citrate buffer (pH 6)
Blocking	Bloxall SP-60002, NHS S-20002, 1:20
Dilution and incubation parameters	1:75, 4 °C, overnight
Secondary antibody used	HaM IgG BA-20002, 1:200
Laminin
Immunoglobuline isotype/clone status	IgG2a/mouse, monoclonal
Catalog no./provider	2E8/DSHB1
Demasking of antigen	N/A
Blocking	3% H2O2 in methanol, NHS S-20002, 1:20
Dilution and incubation parameters	1:5, room temperature, 30 min
Secondary antibody used	HaM IgG BA-20002, 1:200
MMP-2
Immunoglobuline isotype/clone status	IgG/rabbit, polyclonal
Catalog no./provider	97779/Abcam (Cambridge, MA)
Demasking of antigen	N/A
Blocking	Bloxall SP-60002, NGS S-10002, 1:20
Dilution and incubation parameters	1:100, 4 °C, overnight
Secondary antibody used	GaR IgG BA-10002, 1:200
MMP-9
Immunoglobuline isotype/clone status	IgG/rabbit, polyclonal
Catalog no./provider	38898/Abcam
Provider	Abcam
Demasking of antigen	N/A
Blocking	Bloxall SP-60002, NGS S-10002, 1:20
Dilution and incubation parameters	1:100, 4 °C, overnight
Secondary antibody used	GaR IgG BA-10002, 1:200
Procollagen 1
Immunoglobuline isotype/clone status	IgG1/mouse, monoclonal
Catalog no./provider	SP1.D8/DSHB1
Demasking of antigen	Boiling in citrate buffer (pH 6)
Blocking	Bloxall SP-60002, NHS S-20122, 2.5%
Dilution and incubation parameters	1:10, 4 °C, overnight
Secondary antibody used	HaM IgG BA-20002, 1:200
Tenomodulin
Immunoglobuline isotype/clone status	IgG/rabbit, polyclonal
Catalog no./provider	203676/Abcam
Demasking of antigen	N/A
Blocking	3%H2O2 in Methanol, NHS S-20122, 2.5%
Dilution and incubation parameters	1:500, room temperature, 30 min
Secondary antibody used	HaR IgG BA-11002, 1:200
Type I collagen
Immunoglobuline isotype/clone status	IgG1/mouse, monoclonal
Catalog no./provider	C2456/Sigma-Aldrich (St. Louis, MO, United States)
Demasking of antigen	Protease XIV and Hyaluronidase
Blocking	Bloxall SP-60002, NHS S-20002, 1:20
Dilution and incubation parameters	1:2000, room temperature, 30 min
Secondary antibody used	HaM IgG BA-20002, 1:200
Type II collagen
Immunoglobuline isotype/clone status	MIgG2A kappa light chain/mouse, monoclonal
Catalog no./provider	CIIC1/DSHBa
Demasking of antigen	Protease XIV and Hyaluronidase
Blocking	Bloxall SP-60002, NHS S-20002, 1:20
Dilution and incubation parameters	1:6, room temperature, 30 min
Secondary antibody used	HaM IgG BA-20002, 1:200
Type III collagen
Immunoglobuline isotype/clone status	FH-7AIgG1/mouse, monoclonal
Catalog no./provider	C 7805/Sigma-Aldrich (St. Louis, MO, United States)
Demasking of antigen	ProteaseXIV and Hyaluronidase
Blocking
Dilution and incubation parameters	1:4000, room temperature, 30 min
Secondary antibody used	HaM IgG BA-20002, 1:200
Type IV collagen
Immunoglobuline isotype/clone status	MIgG1 kappa light chain/mouse, monoclonal
Catalog no./provider	M3F7/DSHBa
Demasking of antigen	N/A
Blocking	Bloxall SP-60002
Dilution and incubation parameters	1:5, room temperature, 30 min
Secondary antibody used	HaM IgG BA-20002, 1:200

¹The antibodies 12/21/1-C-6 (developed by Dr. Caterson B), 2E8 (developed by Dr. Engvall ES), CIIC1 (developed by Drs Holmdahl R and Rubin K) and SP1.D8 and M3F7 (developed by Dr. Furthmayr H) were obtained from the Developmental Studies Hybridoma Bank (DSHB), created by the Eunice Kennedy Shriver National Institute of Child Health and Human Development of the National Institutes of Health of the United States, and maintained at The University of Iowa, Department of Biology, Iowa City, IA, United States.

²Provider: Vector Laboratories (Burlingame, CA, United States). BSA: Bovine serum albumin; GaM: Goat anti-mouse; GaR: Goat anti-rabbit; HaM: Horse anti-mouse; HaR: Horse anti-rabbit; IgG: Immunoglobulin G; N/A: Not applicable; NHS: Normal horse serum; NGS: Normal goat serum; PBS: Phosphate-buffered saline.