rev: 5/15/2008
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Cytokines, Growth Factors & Chemokines:ANTIBODIES
(anti-Human and
others as indicated)
RDI Division of Fitzgerald Industries Intl offers
a wide line of recombinant growth factors, cytokines and chemokines and
antibodies. Since no one antibody works best for all applications
(neutralization, blotting, ELISA, etc), we offer many different types of
antibodies to help solve this problem. Please inquire for other applications or
types of antibodies not listed below.
Chemokine Receptor polyclonals
-see also anti-Goat secondary antibodies
see sample WB and histo protocols below
$593.25/vial email for specs not linked (in
process)
|
Specificity |
Host |
Size |
Catalog# |
|
Human CCR-1 |
rabbit |
100ul |
|
|
Mouse CCR4 |
rabbit |
100ul |
RDI-MCCR4abrX |
|
Human CCR-5 |
Goat |
200ug |
|
|
Human CXCR-4 |
Goat |
200ug |
|
|
Mouse CXCR-4 |
Goat |
200ug |
RDI-MCXCR4abGX |
|
Human CCR4 |
Goat |
200ug |
|
|
Human CCR2 |
Goat |
200ug |
|
|
Human CCR3 |
Goat |
200ug |
|
|
Human CCR8 n term |
Goat |
200ug |
RDI-CCR8NabGX |
|
Human CCR8 2nd extracellular domain |
Goat |
200ug |
RDI-CCR8XabgX |
|
Mouse CCR4 N term |
Goat |
200ug |
|
|
Mouse CCR8 extracellular domain |
Goat |
200ug |
RDI-MCCR8XabgX |
|
Human CCR9 extracellular domain |
Goat |
200ug |
|
|
Mouse CCR9 extracellular domain |
Goat |
200ug |
|
|
Human CCR-10 extracellular domain |
Goat |
200ug |
|
|
Mouse CCR-10 extracellullar domain |
Goat |
200ug |
|
|
Human CCR-11 ext |
Goat |
200ug |
|
|
Human CCR-6 extracellular domain |
Goat |
200ug |
|
|
Human CCR-D6 |
Goat |
200ug |
RDI-CCRD6abgX |
|
Mouse CCR-7 extracellular domain |
Goat |
100ug |
RDI-MCCR7XabgX |
|
Mouse CCR-D6 |
Goat |
200ug |
RDI-MCCRD6abGX |
|
Human Toll Receptor 2 ext domain |
Goat |
200ug |
RDI-TOLLR2XabgX |
|
Human Toll Receptor 5 ext domain |
Goat |
200ug |
RDI-TOLL5RXabgX |
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Rabbit anti-Human CCR-1 F(ab')2
cat# RDI-CCR1abRX $593.25
Presentation: 100ul of peptide affinity purified F(ab')2 IgG in PBS with 1 mg/ml BSA and 0.1% sodium azide.
host: rabbit IgG
Antigen: Synthetic Peptide
Sequence of Peptide NH2-TEDYDTTTEFDYGDATPC
Specificity: Peptide sequence shares less than 50% amino acid identity to other Chemokine receptors in this region. Since the peptide sequence shares less than 50% amino acid identity to other chemokine receptors in this region, this antibody only recognizes human receptor.
Storage: Freeze at -80 °C. Stable for at least 3 years when stored at -80 °C. Avoid freeze/thaw cycles.
Application: (Working Dilutions)
Western Blotting and Immunohistochemistry on Acetone fixed cells.
ELISA/peptide : 1:50,000
Western Blot: 1:1000
Immunocytochemistry: 1: 300
Immunohistochemistry: 1: 300
Quality Control Test for Each Lot: Western Blot on membranes from human peripheral blood leukocytes or human T-cell line Jurkatt.
References: Neote K, DiGregorio D, Mark J. Y.,Horuk. R.and Schall T.J. 1993. "Molecular Cloning, Functional Expression, and Signaling Characteristics of a C-C Chemokine Receptor." Cell. 72, 415-425.
Su S.B., Mukaida. N., Nomura. H and Matsushima. K. 1996. "Preparation of specific polyclonal antibodies to a C-C chemokine receptor, CCR1, and determination of CCR1expression on various types of leukocytes."
J Leukoc. Biol 60(5), 658-666.
For research Use Only
Goat anti-human CCR2 (affinity purified) Antiserum (affinity purified) to C-C Chemokine Receptor 2 (human)
Cat# RDI-CCR2abGX $562.00
Presentation: 200ug of peptide affinity purified IgG in 0.1ml (2mg/ml) 10mM KHPO4, 140mM NaCl, pH7.2 with 1 mg/ml BSA and 0.1% sodium azide.
Host: Goat IgG
Antigen: Synthetic peptide CGEEVTTFFDYDYGAPCHKFDVKQ corresponding to the N-terminal extracellular domain of human CCR2.
Specificity: Peptide sequence is < 50 % identical to other chemokine receptors in this region. Cross reactivity to mouse CCR2 not tested.
Applications: ELISA 1:60000
Immunoblotting >1:1000
Immunocytochemistry >1:400
Immunohistochemistry >1:200
pos control spleen cells
Flowcytometry N. D
Storage: Freeze at -80 °C. Stable for at least 3 years when stored at -80 °C. Avoid freeze/thaw cycles.
Literature References: [1] Charo,I.F., Myers,S.J., Herman,A., Franci,C., Connolly,A.J. and Coughlin,S.R. Molecular cloning and functional expression of two monocyte chemoattractant protein 1 receptors reveals alternate splicing of the carboxyl-terminal tails. Proc. Natl. Acad. Sci. U.S.A. 91, 2752-2756 (1994)
For research Use Only-not for use in or on humans or animals
Goat anti-human CCR3 affinity purified
cat# RDI-CCR3abGX $562.00
Presentation: 200ug of peptide affinity purified IgG in 0.1ml (2mg/ml) 10mM KHPO4, 140mM NaCl, pH7.2 with 1 mg/ml BSA and 0.1% sodium azide.
Immunogen: 28 amino acid synthetic peptide CVETFGTTSYYDDVGLLCEKADTRALMA
Specificity: Peptide sequence is < 50 % identical to other human chemokine receptors in this region. CCR3 is the receptor for eotaxin and a novel CC Chemokine, leukotactin-1. CCR3 is expressed on eosinophils, TH2 cell subsets, basophils, and mast cells, and plays a key role in allergic diseases.
Applications: ELISA to peptide 1:60000
Immunoblotting 1:1000
Immunocytochemistry 1:300
Immunohistochemistry 1:300
Positve Control: human peripheral blood cells.
Storage/Stability: Freeze at -80 °C. Stable for at least
3 years when stored at -80 °C. Avoid freeze/thaw cycles.
Literature References: -Auger, G.A., et al. 2002. Eur. J. Immunol. 32, 1052;
-Humbles, A.A., et al. 2002. Proc. Natl. Acad.Sci. USA 99, 1479
-
Background: Human CCR3 is a receptor for CC chemokine eotaxin-1 and oetaxin-2. CCR3 is constitutively expressed on eosinophils, small intestine and colon. The human CCR3 protein contains 356 amino acid. Antibody to human CCR3 receptor was generated to a synthetic peptide derived from the amino terminal extracellular domain of the human CCR3 receptor. Goat anti-CCR3 IgG antibody binds to human eosinophils at a dilution of > 1.500.
For Research Use Only
Goat anti-human CCR4 (affinity purified) Antiserum (affinity purified) to C-C Chemokine Receptor 4 (human)
Cat# RDI-CCR4abgX $562.00
Presentation: 200ug of peptide affinity purified IgG in PBS at 1.0mg/ml with
1 mg/ml BSA and 0.1% sodium azide.
Host: Goat
Immunogen: 28 amino acid synthetic peptide L D E S I Y S N Y Y L Y E S
I P K P C T K E G I K A F G E.
derived from the amino terminal extracellular domain of the human CCR4 receptor.
Specificity: Peptide sequence is <50% identical to other Chemokine receptors in this region.
Applications:
ELISA >1: 100000
Western Blot >1: 1000
Immunocytochemistry (acetone fixed cells) >1: 400
Immunohistochemistry (paraffin sections) >1: 200
Not tested for Flowcytometry
Introduction: Human CCR4 is a receptor for macrophage inflammatory protein 1 alpha and Rantes and is expressed on human basophils and 20% of the human adult peripheral blood CD4+ cells. The human CCR4 protein contains 360 amino acids that is 85% identical to mouse CCR4 receptor and contains three glycosylation sites. human CCR4 transcripts are expressed in the thymus and peripheral blood leukocytes.
Storage: Freeze at -80 °C. Stable for at least 3 years when stored at -80 °C. Avoid freeze/thaw cycles.
Literature References:
Receptor cDNA from a Human Basophilic Cell Line. J Biol. Chem. vol. 270 :33: 19495-19500.
For research Use Only
Goat anti-mouse CCR4 N-Terminal Antiserum (affinity purified) to C-C Chemokine Receptor 4 (mouse).
cat# RDI-MCCR4abGX $562.00
Presentation: 200ug peptide affinity purified GOAT IgG in PBS with 1 mg/ml BSA and buffers containing 0.1% sodium azide.
Host: Goat
Antigen: Synthetic peptide DTTQDETVYNSYYFYESMPC corresponding to aa 8-26 of mouse CCR4. This seequence corresponds to amino acid residues 8 - 26 of mouse CC chemokine receptor CCR4.
Specificity: Peptide sequence is < 50 % identical to other human chemokine receptors in this region. The antiserum recognizes mouse CCR4. No cross-reactivity to human CCR4. CCR4 is selectively expressed on Th2-type T cells and as been shown to be responsible for Th2-dominant immune responses. Its ligand is the thymus and activation-regulated chemokine TARC No cross-reactivity to human CCR4.
Applications: ELISA >1:50000
Immunoblotting >1:1000
Immunobytochemistry >1:500 (mouse spleen)
Flow Cytometry 1-3ug/10^6 cells/50-100ul
Storage: Freeze at -80 °C. Stable for at least 3 years when stored at -80 °C. Avoid freeze/thaw cycles.
Literature
References: Molecular cloning of mouse CC CKR-4 and high affinity binding of chemokines to murine and human CC CKR-4: A.J. Hoogewerf, D.Black, A.F. Proudfoot, T.N.C. Wells, and C.A. Power; BBRC 218, 337 (1996)
For Research Use Only
Goat anti-human CCR5 (affinity purified) Antiserum (affinity purified) to C-C Chemokine Receptor 5 (human)
cat# RDI-CCR5NabgX $562.00
Presentation: 200ug of peptide affinity purified IgG in PBS with 1 mg/ml BSA and 0.1% sodium azide.
Antigen: Synthetic peptide (NH2-YQVSSPIYDINYYTSEPCQKINVKQIAA) corresponding to the N-terminal sequence of human CCR5.
Specificity: Binds to CCR 5 receptor on human peripheral blood leukocytes as determined by immunocytochemistry.
Applications:
ELISA 1:50000
Western Blot 1:1000
Immunocytochemistry: 1: 300 (pos control human peripheral blood)
Immunohistochemistry 1: 300 (human spleen)
Background: The HIV receptor is composed of two molecules, the CD4 binding receptor and a co-receptor. The chemokine receptor CCR5 is one of the co-receptors used by primary isolates of HIV-1. Extracellular domains of CCR5 are important for HIV entry into the target cells.
Storage/Stability: Freeze at -80 °C. Stable for at least 3 years when stored at -80 °C. Avoid freeze/thaw cycles.
References:
1. A RANTES, MIP-1a, MIP-1b receptor as fusion cofactor for macrophage-tropic HIV-1: Alkhatib et al.; Science 272, 1955 (1996)
2. Muliple Extracellular Domains of CCR-5 contribute to human immunodeficiency virus type 1 entry and fusion: J.Virol. 71, 5003 (1997)
Goat anti-mouse CCR4 N-Terminal Antiserum (affinity purified) to C-C Chemokine Receptor 4 (mouse).
cat# RDI-MCCR4abGX $562.00
Presentation: 200ug peptide affinity purified GOAT IgG in PBS with 1 mg/ml BSA and buffers containing 0.1% sodium azide.
Host: Goat
Antigen: Synthetic peptide DTTQDETVYNSYYFYESMPC corresponding to aa 8-26 of
mouse CCR4. This seequence corresponds to amino acid residues 8 - 26 of mouse
CC chemokine receptor CCR4.
Specificity: Peptide sequence is < 50 % identical to other human chemokine receptors in this region. The antiserum recognizes mouse CCR4. No cross-reactivity to human CCR4. CCR4 is selectively expressed on Th2-type T cells and has been shown to be responsible for Th2-dominant immune responses. Its ligand is the thymus and activation-regulated chemokine TARC No cross-reactivity to human CCR4.
Applications: ELISA >1:50000
Immunoblotting >1:1000
Immunobytochemistry >1:500 (mouse spleen)
Flow Cytometry 1-3ug/10^6 cells/50-100ul
Storage: Freeze at -80 °C. Stable for at least 3 years when stored at -80 °C. Avoid freeze/thaw cycles.
Literature References: Molecular cloning of mouse CC CKR-4 and high affinity binding of chemokines to murine and human CC CKR-4: A.J. Hoogewerf, D.Black, A.F. Proudfoot, T.N.C. Wells, and C.A. Power; BBRC 218, 337 (1996)
For Research Use Only
Goat anti- Human CxCR4 (affinity purified) N-terminal - extracellular domain
Cat# RDI-CXCR4abGX $562.00
Presentation: 200ug of peptide affinity purified IgG in PBS at 1.0mg/ml with 1 mg/ml BSA and 0.1% sodium azide. Antibody in formulation without azide for neutralization studies is also available.
Host: Goat
Antigen: TEEMGSGDYDSMKEPCFREENANFNKC corresponding to N-terminal extracellular
domain of human CxCR4 receptor.
Specificity: Binds to CXCR 4 receptor on human spleen leukocytes as determined by immunocytochemistry.
Applications: ELISA: 1:50,000
Western Blotting titer 1:1000
Immunocytochemistry 1:300 (human spleen)
PARAFFIN SECTIONS
Storage: Freeze at -80 °C. Stable for at least 3 years when stored at -80 °C. Avoid freeze/thaw cycles.
References:
1. Bleul, C.C etal 1996. The lymphocyte chemoattractant, SDF-1 is a ligand for LESTR/fusin and blocks HIV-1 entry. Nature 382 : 829-833.
2. Loetscher M. etal. 1994. Cloning of a human seven transmembrane domain receptor, LESTR, that is highly expressed in leukocytes. J Biol. Chem. 269, N 1: 232-7
For research Use Only
Goat anti- mouse to CxCR4 (affinity purified) N-terminal - extracellular domain
Cat# RDI-MCXCR4abGX
Presentation: 200ug of peptide affinity purified IgG in PBS at 1.0mg/ml with 1 mg/ml BSA and 0.1% sodium azide. Antibody in formulation without azide for neutralization studies is also available.
Host: Goat
Antigen: NH2-YSEEVGSGDYDSNKEPCFRDENVHFNR
Specificity: Binds to CXCR 4 receptor on mouse spleen leukocytes as determined
by immunocytochemistry.
Applications:
ELISA: 1:50,000
Western Blotting titer 1:1000
Immunocytochemistry 1:300 (mouse spleen)
Storage: Freeze at -80 °C. Stable for at least 3 years when stored at -80 °C. Avoid freeze/thaw cycles.
References: 1. Bieniasz et.al. 1997. Murine CxCR-4 is a functional coreceptor for T-cell tropic and dual tropic strains of human immunodeficiency virus type 1. J Virol. 71: 7097.
2. Heesen, M. etal. 1996. Cloning of the mouse fusin gene. homologue to a human HIV-1 cofactor. J Immunol. 157: 5455.
For research Use Only see also other anti-cxcr4 ab (fusin)
Goat anti-Human CCR-6 Extracellular Domain Human
cat# RDI-CCR6XabGX $562.00
Presentation: 200ug of peptide affinity purified IgG in 0.1ml (2mg/ml) 10mM KHPO4, 140mM NaCl, pH7.2 with 1 mg/ml BSA and 0.1% sodium azide.
Host: Goat IgG
Antigen: Synthetic peptide YFVSVNTSYYSVDSEMLLCSLQEVRQFSR corresponding to a.a. 18-46 of human beta chemokine receptor D6.
Specificity: Peptide sequence is < 50 % identical to other human chemokine receptors in this region. CCR6 is a G-protein-coupled receptor that binds MIP-3a. It is expressed on T cells, B cells, and subsets of dendritic cells, and has been implicated in the adaptive immune response. The antiserum recognizes human CCR6.
Applications: ELISA 1: 75000
Western blot not tested
Immunocytochemistry 1:300
Immunohistochemistry 1:250 (pos control paraffin section human spleen)
Flowcytometry 1-3ul/million cells
Quality Control: Immunocytochemistry on human spleen or peripheral blood.
Storage: Freeze at -80 °C. Stable for at least 3 years when stored at -80 °C. Avoid freeze/thaw cycles.
References: Baba, M. etal, Identification of CCR6, the specific receptor for a Novel Lymphocyte directed CC Chemokine LARC. J. Biol. Chem. 272 (23), 14893-14898 (1997)
For Research Use Only
Goat anti-Mouse C-C Chemok. Receptor-7 Ext.Cell.Dom
cat# RDI-MCCR7XabgX $562.00/100ug
Presentation: 100ug of peptide affinity purified IgG antibody in PBS with 1 mg/ml BSA and 0.1 % sodium azide.
Antigen: Synthetic peptide corresponding to aa 2-22 of mouse CCR7.
Specificity: Peptide sequence is < 50 % identical to other mouse chemokine receptors in this region.
Applications:
ELISA 1:100000
Western Blot 1:1000
Immunocytochemistry 1:500 (mouse spleen)
Immunohistochemistry 1:250
Flowcytometry not tested
Storage: Freeze at -80 °C. Stable for at least 3 years when stored at -80 °C. Avoid freeze/thaw cycles.
Literature References: Cloning of human and mouse EBI1, a lymphoid-specific G-protein-coupled receptor encoded on human chromosome 17q12-q21.2. Genomics 23 (3), 643-650 (1994)
For In Vitro Research Use Only
Goat anti-Mouse C-C Chemok. Receptor-8 Ext.Cell.Dom
cat# RDI-MCCR8XabgX $562.00/100ug
Presentation: 200ug of peptide affinity purified IgG antibody in 0.1ml (2mg/ml) PBS (1mM KHPO4, 140mM NaCl pH7.2) with 1mg/ml BSA and 0.1 % sodium azide.
Antigen: Synthetic peptide PNVTMTDYYPDFFTAPCDAEFLLRGSM corresponding to a 7-33 of mouse CCR8
Specificity: Peptide sequence is < 50 % identical to other mouse chemokine receptors in this region. The antiserum recognizes mouse CCR8. Not tested for cross-reactivity to human CCR8.
Applications:
ELISA 1:100000
Western Blot 1:1000
Immunocytochemistry 1:400 (mouse spleen)
Immunohistochemistry 1:400
Flowcytometry not tested
Positive Control: mouse spleen (paraffin sections)
Storage: Freeze at -80 °C. Stable for at least 3 years when stored at -80 °C. Avoid freeze/thaw cycles.
Literature
References: 1. Zingoni, A. et. al. The chemokine receptor CCR8 is preferentially expressed in Th2 but not Th1 cells J. Immunol. 161 (2), 547-551 (1998)
For Research Use Only
Goat anti-human C-C chemokine Receptor-9, Ext.Cell.Dom
cat# RDI-CCR9XabgX $562.00
Presentation: 200ug of peptide affinity purified IgG in 0.1ml (2mg/ml) 10mM KHPO4, 140mM NaCl, pH7.2 with 1 mg/ml BSA and 0.1% sodium azide.
Host: Goat IgG
Antigen: Synthetic peptide IPNMADDYGSESTSSMEDYVNFNFTDFYC corresponding to aa
10-37 of human CCR9.
Specificity: Peptide sequence is < 50 % identical to other human chemokine receptors in this region. The antiserum recognizes human CCR9 and is likely to cross react to mouse CCR9 with approximately 50% less titer .
Applications:
ELISA (human CCR peptide)1: 75000
Western Blotting 1:1000
Immunocytochemistry 1:300
Immunohistochemistry 1:250
Flowcytometry-not tested
Quality Control: Immunocytochemistry on human thymus, spleen or MOLT4 cell line
Storage: Freeze at -80 °C. Stable for at least 3 years when stored at -80 °C. Avoid freeze/thaw cycles.
References: [1] Zaballos,A.,. et. al. identification of the orphan chemokine receptor GPR-9-6 as CCR9, the receptor for the chemokine TECK. J. Immunol. 162 (10), 5671-5675 (1999)
[2] Yu, et. al. "CCR9A and CCR9B: Two Receptors for the Chemokine CCL25/TECK/Ckbeta-15 That Differ in Their Sensitivities to Ligand. J Immunol. 164, 1293-1305, 2000
For Research Use Only
Goat anti-mouse CCR9
cat# RDI-MCCR9XabGX $562.00
Presentation: 200ug of peptide affinity purified IgG in 0.1ml (2mg/ml) 10mM KHPO4, 140mM NaCl, pH7.2 with 1 mg/ml BSA and 0.1% sodium azide.
Host: Goat IgG
Antigen: Synthetic peptide IPGMFDDFSYDSTASTDDYMNLNFSSFF corresponding to aa 10-37 of mouse CCR9.
Specificity: Peptide sequence is < 50 % identical to other mouse chemokine receptors in this region. The antiserum recognizes mouse CCR9 and is likely to cross-react to human CCR9 with approximately 50% less titer.
Applications:
ELISA 1: 100000
Western blot 1:1000
Immunocytochemistry 1:500
Immonuhistochemistry 1:250
Flowcytometry 1:200
Quality Control: Immunocytochemistry on mouse thymus or lymphnodes.
Storage: Freeze at -80 °C. Stable for at least 3 years when stored at -80 °C. Avoid freeze/thaw cycles.
References: Zaballos,A.,. et. al. identification of the orphan chemokine receptor GPR-9-6 as CCR9, the receptor for the chemokine TECK. J. Immunol. 162 (10), 5671-5675 (1999)
For Research Use Only
Goat anti-mouse CCR10
cat# RDI-MCCR10XabGX $562.00
Presentation: 200ug of peptide affinity purified IgG in 0.1ml (2mg/ml) 10mM KHPO4, 140mM NaCl, pH7.2 with 1 mg/ml BSA and 0.1% sodium azide.
Host: Goat IgG
Antigen: Synthetic peptide KPTEQVSWGLYSGYDEEAYSVGPLPELC corresponding to aa 4-31 of mouse CCR10.
Specificity: Peptide sequence is < 50 % identical to other mouse chemokine receptors in this region. CCR10 is the receptor for the skin-associated CC chemokine CCL27 (CTACK) as well as a novel chemokine CCL28 that displays chemotactic activity for resting CD4+ and CD8+ T cells. The antiserum recognizes mouse CCR10 and has minimal cross-reactivity to human CCR10.
Applications: ELISA 1: 100000
Western blot 1:1000
Immunocytochemistry 1:500
Immonohistochemistry 1:250
Flowcytometry not tested
Quality Control: Immunocytochemistry on mouse lymphnodes or small intestine
Storage: Freeze at -80 °C. Stable for at least 3 years when stored at -80 °C. Avoid freeze/thaw cycles.
References: 1] Zaballos,A., et.al. Cutting edge: identification of the orphan chemokine receptor GPR-9-6 as CCR9, the receptor for the chemokine TECK. J. Immunol. 162 (10), 5671-5675 (1999).
[2] Wang, W., et al. 2000. J. Biol. Chem. 275, 22313.
For Research Use Only
cat# RDI-CCR10XabGX $562.00
Presentation: 200ug of peptide affinity purified IgG in 0.1ml (2mg/ml) 10mM KHPO4, 140mM NaCl, pH7.2 with 1 mg/ml BSA and 0.1% sodium azide.
Host: Goat IgG
Antigen: Synthetic peptide VSWGHYSGDEEDAYSAEPLPELC corresponding to aa 9-31 of human CCR10.
Specificity: Peptide sequence is < 50 % identical to other mouse chemokine receptors in this region. CCR10 is the receptor for the skin-associated CC chemokine CCL27 (CTACK) as well as a novel chemokine CCL28 that displays chemotactic activity for resting CD4+ and CD8+ T cells. The antiserum recognizes human CCR10 (not tested with mouse CCR10)
Applications: ELISA 1: 75K
Western blot 1:1000
Immunocytochemistry 1:250
Immunohistochemistry 1:250
Flowcytometry not tested
Quality Control: Immunocytochemistry on paraffin section of human spleen
Storage: Freeze at -70 °C. Stable for at least 3 years when stored at -70 °C. Avoid freeze/thaw cycles.
References: 1] Zaballos,A., et.al. Cutting edge: identification of the orphan chemokine receptor GPR-9-6 as CCR9, the receptor for the chemokine TECK. J. Immunol. 162 (10), 5671-5675 (1999).
[2] Wang, W., et al. 2000. J. Biol. Chem. 275, 22313.
For Research Use Only
Goat anti-human C-C chemokine Receptor-11, Ext.Cell.Dom (Human/Mouse)
Cat #: RDI-CCR11XabGX $562.00
Presentation: 200ug of peptide affinity purified IgG in 0.1ml (2mg/ml) 10mM KHPO4, 140mM NaCl, pH7.2 with 1 mg/ml BSA and 0.1% sodium azide.
Host: Goat IgG
Antigen: Synthetic peptide EQNQSTDYYYEENEMNGTYDYSQYELIC corresponding to aa 4-31 of human CCR11.
Specificity: Peptide sequence is < 50 % identical to other human chemokine receptors in this region. Will cross-react to mouse CCR11.
Applications: ELISA 1:75000
Western Blot 1:1000
Immunocytochemistry 1:500
Immunohistochemistry 1:250 (human lung tissue)
Flowcytometry N. D.
Storage: Freeze at -80 °C. Stable for at least 3 years when stored at -80 °C. Avoid freeze/thaw cycles.
Literature
References: 1] Schweickart,V.L., etal. CCR11 is a functional receptor for the monocyte chemoattractant protein family of chemokines. J. Biol. Chem. 275 (13), 9550-9556 (2000)
For Research Use Only-not for use in or on humans or animals
Immunohistochemistry with antibody to Human CCR11 on paraffin section of mouse Lung
Immunohistochemistry on Paraffin sections:
Paraffin sections of tissues fixed in 10% formalin were used for QC of the immunodetect antibodies. De-wax the sections and block the endogenous peroxide by incubating the sections with 3% H2O2 for 15 minutes at room temperature (RT). Wash the sections with PBS and incubate with 2% normal rabbit serum at room temperature to block the non-specific binding sites. Wash the section with PBS and incubate with primary antibody (anti-chemokine receptor) at recommended dilution for 60 minutes at RT. Wash the slides and incubate at RT with biotinylated rabbit anti-goat IgG H+L (Vector Labs) at a 1:200 dilution for 30 minutes. Wash the slides and incubate with HRP-Streptavidin (Vector Labs) 1:400 dilution for 30 minutes. Wash the slides and incubate with DAB (Sigma) for 5-7 minutes at RT. Wash slides with PBS and counter stain with hematoxylin (DAKO).
Immunocytochemistry & Immunohistochemistry
Immunocytochemistry on cytospun cells or frozen tissues section:
Fix the cells cytospun on slides or frozen tissue section in cold acetone (acetone prechilled to -20C) for 2-5 minutes. Let the slides dry and then block endogenous peroxide activity by treating the slides with peroxide blocker (3% hydrogen peroxide) for 10 minutes. If the secondary antibody is from rabbit then cells should be incubated with 2% normal rabbit serum in PBS. Wash the slide in PBS x 5 then incubate with the antibody at appropriate dilution in PBS containing Ig free 2% BSA or 2% milk for 30-45 minutes at room temperature, wash 5 times with PBS and then incubate with second HRP-conjugated antibody at a dilution optimized previously for 30 minutes at room temperature (If using primary cells from lymphoid organs containing B cells as well the secondary antibody must be specifies specific to minimize staining of B cells by secondary antibody alone). Wash 5 times with PBS and develop the stain by adding the DAB or AEC and counter stain with hematoxylene.
Western Blotting
SDS-PAGE: A NOVEX gel system or any other gel system based on Laemmli's SDS-PAGE system is recommended. If protein of interest is expressed at very low levels in the cells and/or only a very small percentage of cells in the tissue express the desired protein, then a higher amount of total tissue lysate or crude membrane lysate in the range of 100-150 ug of protein per lane is recommended. SDS-PAGE gels used should contain appropriate percent acrylamide to provide the best separation of proteins in the molecular weight range of the desired protein to be detected. We strongly recommend separating proteins on SDS-PAGE gels at 15-20 mA constant current when 100-150 ug of protein is loaded in each lane to ensure clear separation. SDS-PAGE can be run overnight at 5-7 mA constant current and next day in the morning the current should be increased to 20 mA until the dye front reaches the bottom of the gel. This will tighten the bands in the gel.
Transfer of Proteins to Nitrocellulose: When transferring hydrophobic integral cell membrane proteins Tris-Glycine buffer containing 1 x SDS running buffer with 10 to 20% methanol should be used to facilitate transfer of proteins like CCRs that have 7 hydrophobic domains. Tris-Glycine buffer with 10-20% methanol is adequate to transfer most secreted proteins. Transfer the proteins following the instructions provided by the manufacturer of the transfer apparatus. For the semi-dry transfer system we used, protein transfer was done at 150 mA constant current for 60-90 minutes.
Immunodetection: After the transfer, air dry the Nitrocellulose membrane on a paper towel. Rinse the membrane in Tris buffered saline with 0.1% Tween 20 (20 mM Tris, 150 mM NaCl pH 7.5-8.0) for 5 minutes. Block the membrane in 5% milk in TBS-Tween for a minimum of 1 hour. Incubate with anti-chemokine receptor antibody diluted in 1-2% milk in TBS-Tween at room temperature for 2 hours or overnight at 4 C. Wash the membrane x 5, 5 minutes each with TBS-Tween containing 300 mM NaCl (Note TBS-Tween indicated above contains 150mM NaCl. Higher salt is used to facilitate removal of non-specifically bound antibody). Incubate with HRP conjugated secondary antibody at appropriate dilution in 1-2% milk in TBS-Tween containing 150 mM NaCl for a minimum of 1 hour. If using B cells the secondary antibody should be species specific to minimize detection of Ig expressed by B cells by secondary antibody. Wash the membrane x 5, 5 minutes each with TBS-Tween containing 300 mM NaCl. Visualize the bands using the ECL system of Amersham-Pharmacia.
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