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Anti Human CD49d Antibody, clone HP2/1

Mouse Anti-Human Monoclonal Antibody

     
  • IHC - Anti Human CD49d Antibody, clone HP2/1  ABD12839
    Published customer image:Mouse anti Human CD49d antibody, clone HP2/1 used for immunoprecipitationImage caption:Several anti-tetraspanin antibodies co-precipitate ß1 integrins from HEL cells solubilised in Brij-97. Precipitates were prepared from HEL cells solubilised in different detergents in the presence of Mn2+. CD53, MEM-53; CD63, MEM-259; CD81, 454720; CD82, TS82b; CD151, IIG5a; a4, HP2/1; aL, TS1/22; aIIb, PAB-1. Precipitates were run on 7.5% non-reduced gels.From: Spring FA, Griffiths RE, Mankelow TJ, Agnew C, Parsons SF, et al. Tetraspanins CD81 and CD82 Facilitate a4ß1-Mediated Adhesion of Human Erythroblasts to Vascular Cell Adhesion Molecule-1. PLoS ONE 8(5): e62654.
  • WB - Anti Human CD49d Antibody, clone HP2/1  ABD12839
    Published customer image:Mouse anti Human CD49d antibody, clone HP2/1 used for western blotting and immunoprecipitationImage caption:Tetraspanins CD81, CD82 and CD151 are associated with a4ß1 throughout erythroid maturation and with ß3 in proerythroblasts and basophilic erythroblasts. A. CD81, CD82 and CD151 precipitates from Mn2+-activated proerythroblasts (ProEB, day 5), basophilic (BasoEB, day 8) and polychromatic (PolyEB, day 12) erythroblasts were successively probed with anti-a4, anti-ß1 and anti-ß3 antibodies; tetraspanin controls from each time point are also illustrated. All tetraspanins co-precipitated a4 and ß1 from erythroblasts B. Tetraspanin precipitates from day 6 proerythroblasts solubilised in the presence of EDTA or different cations, and from Mn2+-activated basophilic erythroblasts (BasoEB, day 8) were probed with a mix of antibodies to a5, ß1, ß2 and ß3 integrins while the control samples were probed with the relevant tetraspanin antibodies. For clarity, integrin controls are illustrated for the EDTA blot but were present on all blots. ß1 and ß3 integrins were precipitated well only in the presence of Mn2+. C. CD81 and CD82 precipitates from day 5 proerythroblasts were successively probed with different anti-integrin subunit antibodies and demonstrate co-precipitation of ß1 and ß3 but not a5 or ß2 integrins. D. CD81 and CD82 precipitates from day 6 proerythroblasts and HEL cells solubilised in the presence of EDTA, Ca2++Mg2+ or Mn2+ probed with anti-CD82 and anti-CD81 antibodies. Each tetraspanin co-precipitates the other most strongly in the presence of Mn2+ from proerythroblasts while any cation permits co-precipitation in HEL cells. Integrins were analysed on 7.5% gels, tetraspanins on 12% gels; non-reducing conditions. Unless stated, the following clones were used: CD53, MEM-53; CD63, MEM-259; CD81, 454720; CD82, TS82b; CD151, IIG5a; a4, HP2/1; a5, IIA1; aL, TS1/22; aIIb, PAB-1. All day 5 and 6 cultures comprised 90–95% proerythroblasts; day 8 culture comprised 5% proerythroblasts, 81% basophilic erythroblasts and 14% polychromatic erythroblasts; day 12 culture comprised 41% polychromatic erythroblasts, 15% orthochromatic erythroblasts and 41% reticulocytes. In the day 5 and 6 cultures 15–34% of cells were GPA+ and 28–35% of cells were aIIb+. Day 8 and day 12 cultures had 77% and 97% GPA+ cells, respectively, and 9% and 0% aIIb+ cells, respectively.From: Spring FA, Griffiths RE, Mankelow TJ, Agnew C, Parsons SF, et al. Tetraspanins CD81 and CD82 Facilitate a4ß1-Mediated Adhesion of Human Erythroblasts to Vascular Cell Adhesion Molecule-1. PLoS ONE 8(5): e62654.
  • IP - Anti Human CD49d Antibody, clone HP2/1  ABD12839
    Published customer image:Mouse anti Human CD49d antibody, clone HP2/1 used for immunoprecipitationImage caption:All anti-CD81, anti-CD82 and anti-CD151 clones co-precipitate ß1 and ß3 integrins from normal and leukemic proerythroblasts. A. CD81 clones. B. CD82 clones. C. CD151 clones. ERB, day 6 proerythroblasts. a4, HP2/1; a5, IIA1; aL, TS1/22; aIIb, PAB-1. Integrins in 7.5% NR gels, tetraspanins in 12% non-reduced gels. More ß3 integrins are co-precipitated from HEL cells as they express aIIbß3 and aVß3; proerythroblasts express only aIIbß3.From: Spring FA, Griffiths RE, Mankelow TJ, Agnew C, Parsons SF, et al. Tetraspanins CD81 and CD82 Facilitate a4ß1-Mediated Adhesion of Human Erythroblasts to Vascular Cell Adhesion Molecule-1. PLoS ONE 8(5): e62654.
  • IP - Anti Human CD49d Antibody, clone HP2/1  ABD12839
    Published customer image:Mouse anti Human CD49d antibody, clone HP2/1 used for immunoprecipitationImage caption:All anti-CD81, anti-CD82 and anti-CD151 clones co-precipitate ß1 and ß3 integrins from normal and leukemic proerythroblasts. A. CD81 clones. B. CD82 clones. C. CD151 clones. ERB, day 6 proerythroblasts. a4, HP2/1; a5, IIA1; aL, TS1/22; aIIb, PAB-1. Integrins in 7.5% NR gels, tetraspanins in 12% non-reduced gels. More ß3 integrins are co-precipitated from HEL cells as they express aIIbß3 and aVß3; proerythroblasts express only aIIbß3.From: Spring FA, Griffiths RE, Mankelow TJ, Agnew C, Parsons SF, et al. Tetraspanins CD81 and CD82 Facilitate a4ß1-Mediated Adhesion of Human Erythroblasts to Vascular Cell Adhesion Molecule-1. PLoS ONE 8(5): e62654.
  • WB - Anti Human CD49d Antibody, clone HP2/1  ABD12839
    Published customer image:Mouse anti Human CD49d antibody, clone HP2/1 used for blocking studies.Image caption: Blocking experiments with anti-ß7, anti-a4 and anti-aVß3 antibodies for adhesion to HS-5 stromal cells. Panel A, Blocking with anti-ß7. HMCLs were stimulated with Pam3CSK4 for 24 hours and then treated with anti-ß7 antibody before adhesion to HS5-coated wells. Panel B, Blocking experiments with anti-a4 and anti-aVß3 antibodies for adhesion to HS-5. HMCLs were stimulated with Pam3CSK4 for 24 hours and then treated with anti-a4 and anti-aVß3 antibodies before adhesion to HS5-coated wells. The results are the statistical analyses of data in 3 separate experiments expressed as mean ± SEM, *P
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Product Information
Application
  • Applications Legend:
  • WB=Western Blot
  • IHC=Immunohistochemistry
  • IHC-P=Immunohistochemistry (Paraffin-embedded Sections)
  • IHC-F=Immunohistochemistry (Frozen Sections)
  • IF=Immunofluorescence
  • FC=Flow Cytopmetry
  • IC=Immunochemistry
  • ICC=Immunocytochemistry
  • E=ELISA
  • IP=Immunoprecipitation
  • DB=Dot Blot
  • CHIP=Chromatin Immunoprecipitation
  • FA=Fluorescence Assay
  • IEM=Immunoelectronmicroscopy
  • EIA=Enzyme Immunoassay
IHC-F, FC, IP, Func
Primary Accession P13612
Reactivity Human
Host Mouse
Clonality Monoclonal
Isotype IgG1
Clone Names HP2/1
Calculated MW 114900 Da
Additional Information
Other Species Rat,Rh,B,Pig,Cynomolgus monkey,Ga,Rb,Llama,Hs,Mink,Mustelid,Cat
Purification Purified IgG prepared from ascites or tissue culture supernatant
Immunogen JM leukaemia line.
Shelf Life 18 months from date of despatch.
Gene ID 3676
Other Names Integrin alpha-4, CD49 antigen-like family member D, Integrin alpha-IV, VLA-4 subunit alpha, CD49d, ITGA4, CD49D
Target/Specificity Mouse anti Human CD49d monoclonal antibody, clone HP2/1 recognizes human CD49d also known as integrin alpha-4 or VLA-4 subunit alpha. CD49d is a ~150kDa single pass type 1 transmembrane glycoprotein with seven FG-GAP repeats, characteristic of alpha integrins, in its extracellular domain. CD49d can be proteolytically cleaved to yield framents of 80 and 70kDa (Hemleret al.1987). CD49d associates with either CD29 to form VLA-4 or with Integrin beta-7 to form The Peyer patches-specific homing receptor LPAM-1, involved in the lymphocyte migration and homing to gut-associated lymphoid tissue (Sackstein 2006) through its interaction with MadCam-1, preferentially expressed on Peyer’s patch high endothelial venules and postcapillary venules in lamina propria (Briskinet al.1997).Mouse anti human CD49d, clone HP2/1 binds to both intact and the 80kDa fragment of integrin alpha-4. CD49d is expressed on monocytes, T cells, B cells, thymocytes and Langerhans cells (de Graaf et al.1995).Mouse anti Human CD49d, clone HP2/1 can be used in basic studies of VLA-4 mediated adhesion and its interaction with the VCAM-1 structure and has been demonstrated to inhibit cell binding to soluble VCAM-1 (Welleret al.1991).
Preservative & Stabilisers 0.09%Sodium Azide
Storage Store at +4℃ or at -20℃ if preferred.
PrecautionsAnti Human CD49d Antibody, clone HP2/1 is for research use only and not for use in diagnostic or therapeutic procedures.
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