CD49d antibody | HP2/1

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Anti Human CD49d Antibody, clone HP2/1 thumbnail image 1

Anti Human CD49d Antibody, clone HP2/1 thumbnail image 2

Anti Human CD49d Antibody, clone HP2/1 thumbnail image 3

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Anti Human CD49d Antibody, clone HP2/1 gallery image 1 — **Published customer image:**
Mouse anti Human CD49d antibody, clone HP2/1 used for binding efficiency determination
**Image caption:**
Binding efficiencies (BE) of different α4β7 molecules composed of distinct α4 mutants to monoclonal antibodies against α4, β7 or the α4β7 heterodimer. Binding efficiency is determined by the ratio between the mean fluorescence of antibody binding to each α4 molecule and of the binding in a mock-transfected cell culture (see Materials and Methods for details). Dark gray bars represent binding to the human (wild type) α4 clone, whereas light gray bars are those of binding to the different α4 mutants (as shown in the x-axis). α4 mutants which included substitutions at codon 201 are boxed. A, binding of anti-α4 2b4 antibody. B, binding of anti-α4 HP2/1 antibody. C, BE of different anti-α4 and β7 antibodies to the human α4 and the quintuple α4 mutant (5 aa mut). Bars represent the range of standard errors deduced from triplicate experiments. p-values of Student's t tests are shown above each comparison. NS, non-significant (> 0.05).

From:Darc M, Hait SH, Soares EA, Cicala C, Seuanez HN, et al. (2011) Polymorphisms in the α4 Integrin of Neotropical Primates: Insights for Binding of Natural Ligands and HIV-1 gp120 to the Human α4β7. PLoS ONE 6(9): e24461.

Anti Human CD49d Antibody, clone HP2/1 gallery image 2 — **Published customer image:**
Mouse anti Human CD49d antibody, clone HP2/1 used for western blotting and immunoprecipitation
**Image caption:**
Tetraspanins CD81, CD82 and CD151 are associated with α4β1 throughout erythroid maturation and with β3 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-α4, anti-β1 and anti-β3 antibodies; tetraspanin controls from each time point are also illustrated. All tetraspanins co-precipitated α4 and β1 from erythroblasts B. Tetraspanin precipitates from day 6 proerythroblasts (ProEB) 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 α5, β1, β2 and β3 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. β1 and β3 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 β1 and β3 but not α5 or β2 integrins. D. CD81 (454720) and CD82 (53H5) precipitates from day 6 proerythroblasts (ProEB) and HEL cells (HEL) 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; α4, HP2/1; α5, IIA1; αL, TS1/22; αIIb, 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 αIIb+. Day 8 and day 12 cultures had 77% and 97% GPA+ cells, respectively, and 9% and 0% αIIb+ cells, respectively.

From: Spring FA, Griffiths RE, Mankelow TJ, Agnew C, Parsons SF, et al. (2013) Tetraspanins CD81 and CD82 Facilitate α4β1-Mediated Adhesion of Human Erythroblasts to Vascular Cell Adhesion Molecule-1. PLoS ONE 8(5): e62654.

Anti Human CD49d Antibody, clone HP2/1 gallery image 3 — **Published customer image:**
Mouse anti Human CD49d antibody, clone HP2/1 used for immunoprecipitation
**Image caption:**
Several anti-tetraspanin antibodies co-precipitate β1 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; α4, HP2/1; αL, TS1/22; αIIb, PAB-1. Precipitates were run on 7.5% non-reduced gels.

From: Spring FA, Griffiths RE, Mankelow TJ, Agnew C, Parsons SF, et al. (2013) Tetraspanins CD81 and CD82 Facilitate α4β1-Mediated Adhesion of Human Erythroblasts to Vascular Cell Adhesion Molecule-1. PLoS ONE 8(5): e62654.

Anti Human CD49d Antibody, clone HP2/1 gallery image 4 — **Published customer image:**
Mouse anti Human CD49d antibody, clone HP2/1 used for immunoprecipitation
**Image caption:**
All anti-CD81, anti-CD82 and anti-CD151 clones co-precipitate β1 and β3 integrins from normal and leukemic proerythroblasts. A. CD81 clones. B. CD82 clones. C. CD151 clones. ERB, day 6 proerythroblasts. α4, HP2/1; α5, IIA1; αL, TS1/22; αIIb, PAB-1. Integrins in 7.5% NR gels, tetraspanins in 12% non-reduced gels. More β3 integrins are co-precipitated from HEL cells as they express αIIbβ3 and αVβ3; proerythroblasts express only αIIbβ3.

From: Spring FA, Griffiths RE, Mankelow TJ, Agnew C, Parsons SF, et al. (2013) Tetraspanins CD81 and CD82 Facilitate α4β1-Mediated Adhesion of Human Erythroblasts to Vascular Cell Adhesion Molecule-1. PLoS ONE 8(5): e62654.

Anti Human CD49d Antibody, clone HP2/1 gallery image 5 — **Published customer image:**
Mouse anti Human CD49d antibody, clone HP2/1 used for immunoprecipitation
**Image caption:**
All anti-CD81, anti-CD82 and anti-CD151 clones co-precipitate β1 and β3 integrins from normal and leukemic proerythroblasts. A. CD81 clones. B. CD82 clones. C. CD151 clones. ERB, day 6 proerythroblasts. α4, HP2/1; α5, IIA1; αL, TS1/22; αIIb, PAB-1. Integrins in 7.5% NR gels, tetraspanins in 12% non-reduced gels. More β3 integrins are co-precipitated from HEL cells as they express αIIbβ3 and αVβ3; proerythroblasts express only αIIbβ3.

From: Spring FA, Griffiths RE, Mankelow TJ, Agnew C, Parsons SF, et al. (2013) Tetraspanins CD81 and CD82 Facilitate α4β1-Mediated Adhesion of Human Erythroblasts to Vascular Cell Adhesion Molecule-1. PLoS ONE 8(5): e62654.

Anti Human CD49d Antibody, clone HP2/1 gallery image 6 — **Published customer image:**
Mouse anti Human CD49d antibody, clone HP2/1 used for blocking studies.
**Image caption:**
Blocking experiments with anti-β7, anti-α4 and anti-αVβ3 antibodies for adhesion to HS-5 stromal cells. Panel A, Blocking with anti-β7. HMCLs were stimulated with Pam3CSK4 for 24 hours and then treated with anti-β7 antibody before adhesion to HS5-coated wells. Panel B, Blocking experiments with anti-α4 and anti-αVβ3 antibodies for adhesion to HS-5. HMCLs were stimulated with Pam3CSK4 for 24 hours and then treated with anti-α4 and anti-αVβ3 antibodies before adhesion to HS5-coated wells. The results are the statistical analyses of data in 3 separate experiments expressed as mean ± SEM, *P<0.05, **P<0.01, ***P<0.001.

From: Abdi J, Mutis T, Garssen J, Redegeld FA (2014) Toll-Like Receptor (TLR)-1/2 Triggering of Multiple Myeloma Cells Modulates Their Adhesion to Bone Marrow Stromal Cells and Enhances Bortezomib-Induced Apoptosis. PLoS ONE 9(5): e96608.

Mouse anti Human CD49d

Product Type: Monoclonal Antibody
Clone: HP2/1
Isotype: IgG1

Product Code	Applications	Datasheet	MSDS	Pack Size	List Price	Quantity
MCA697GA	C F FN* IP			0.1 mg
MCA697	C F FN* IP			0.2 mg

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 (Hemler et 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 (Briskin et 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 (Weller et al. 1991).

Product Details

Target Species

Human

Species Cross-Reactivity

Target Species	Cross Reactivity
Rat
Rhesus Monkey
Bovine
Pig
Cynomolgus monkey
Goat
Rabbit
Llama
Horse
Mink
Mustelid
Cat

N.B. Antibody reactivity and working conditions may vary between species.

Product Form

Purified IgG - liquid

Preparation

Purified IgG prepared from ascites or tissue culture supernatant

Buffer Solution

Phosphate buffered saline

Preservative Stabilisers

0.09%

Sodium Azide

Immunogen

JM leukaemia line.

Approx. Protein Concentrations

IgG concentration 1 mg/ml

Fusion Partners

Spleen cells from immunised BALB/c mice were fused with cells of the X63 Ag8.653 myeloma cell line.

Storage Information

Storage: Store at +4^oC or at -20^oC if preferred.

This product should be stored undiluted.

Storage in frost free freezers is not recommended. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
Shelf Life: 18 months from date of despatch.

More Information

UniProt: P13612 Related reagents
Entrez Gene: ITGA4 Related reagents
GO Terms: GO:0007596 blood coagulation; GO:0004872 receptor activity; GO:0008305 integrin complex; GO:0007159 leukocyte cell-cell adhesion; GO:0007229 integrin-mediated signaling pathway; GO:0042802 identical protein binding; GO:0030183 B cell differentiation; GO:0050776 regulation of immune response; GO:0050900 leukocyte migration
Regulatory: For research purposes only

Applications of CD49d antibody

This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.

Application Name	Verified	Min Dilution	Max Dilution
Flow Cytometry			1ug/5 x 10⁵ cells
Functional Assays ¹
Immunohistology - Frozen
Immunoprecipitation

¹This product contains sodium azide, removal by dialysis is recommended prior to use in functional assays. Bio-Rad recommend the use of EQU003 for this purpose.

Where this antibody has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the antibody for use in their own system using appropriate negative/positive controls.

Secondary Antibodies Available

Description	Product Code	Pack Size	Applications
Human anti Mouse IgG1:HRP	HCA036P	0.1 mg	E
Goat anti Mouse IgG (H/L):Alk. Phos. (Multi Species Adsorbed)	STAR117A	0.5 mg	E WB
Goat anti Mouse IgG (H/L):DyLight®488 (Multi Species Adsorbed)	STAR117D488GA	0.1 mg	F IF
Goat anti Mouse IgG (H/L):DyLight®549 (Multi Species Adsorbed)	STAR117D549GA	0.1 mg	F IF WB
Goat anti Mouse IgG (H/L):DyLight®649 (Multi Species Adsorbed)	STAR117D649GA	0.1 mg	F IF
Goat anti Mouse IgG (H/L):DyLight®680 (Multi Species Adsorbed)	STAR117D680GA	0.1 mg	F WB
Goat anti Mouse IgG (H/L):DyLight®800 (Multi Species Adsorbed)	STAR117D800GA	0.1 mg	F IF WB
Goat anti Mouse IgG (H/L):FITC (Multi Species Adsorbed)	STAR117F	0.5 mg	F
Goat anti Mouse IgG (H/L):HRP (Multi Species Adsorbed)	STAR117P	0.5 mg	E WB
Goat anti Mouse IgG (Fc):FITC	STAR120F	1 mg	C F
Goat anti Mouse IgG (Fc):HRP	STAR120P	1 mg	E WB
Rabbit F(ab')2 anti Mouse IgG:RPE	STAR12A	1 ml	F
Rabbit F(ab')2 anti Mouse IgG:HRP (Human Adsorbed)	STAR13B	1 mg	C E P RE WB
Goat anti Mouse IgG:FITC (Rat Adsorbed)	STAR70	0.5 mg	F
Goat anti Mouse IgG:RPE (Rat Adsorbed)	STAR76	1 ml	F
Goat anti Mouse IgG:HRP (Rat Adsorbed)	STAR77	0.5 mg	C E P
Goat anti Mouse IgG/A/M:Alk. Phos.	STAR87A	1 mg	C E WB
Goat anti Mouse IgG/A/M:HRP (Human Adsorbed)	STAR87P	1 mg	E
Rabbit F(ab')2 anti Mouse IgG:Dylight®800	STAR8D800GA	0.1 mg	F IF WB
Rabbit F(ab')2 anti Mouse IgG:FITC	STAR9B	1 mg	F

Negative Isotype Controls Available

Description	Product Code	Pack Size	Applications	List Price	Quantity
Mouse IgG1 Negative Control	MCA928	100 Tests	F

Application Based External Images

Flow Cytometry

Functional Assays

Product Specific References

References for CD49d antibody

Sánchez-Madrid, F. et al. (1986) VLA-3: a novel polypeptide association within the VLA molecular complex: cell distribution and biochemical characterization.
Eur J Immunol. 16 (11): 1343-9.
Weller, P. F. et al. (1991) Human eosinophil adherence to vascular endothelium mediated by binding to vascular cell adhesion molecule 1 and endothelial leukocyte adhesion molecule 1.
Proc Natl Acad Sci U S A. 88: 7430-3.
Mattila, P. et al. (1992) VLA-4 integrin on sarcoma cell lines recognizes endothelial VCAM-1. Differential regulation of the VLA-4 avidity on various sarcoma cell lines.
Int J Cancer. 52 (6): 918-23.
Kumagai, M. et al. (1995) The cross-reactivity of anti-human adhesion mAb with primate and swine cells.
Leucocyte Typing V. Oxford University Press p. 1646-8.
Sopper, S. et al. (1997) Lymphocyte subsets and expression of differentiation markers in blood and lymphoid organs of rhesus monkeys.
Cytometry. 29 (4): 351-62.
Van Vliet, S. S. et al. (1995) Species cross reactivity (human-monkey-pig-bovine) of the adhesion structure section mAB.
Leucocyte Typing V. Oxford University Press p 1607-8.
Meister, R.K. et al. (2007) Progress in the discovery and definition of monoclonal antibodies for use in feline research.
Vet Immunol Immunopathol. 119: 38-46.
Bode, U. et al. (2008) Dendritic cell subsets in lymph nodes are characterized by the specific draining area and influence the phenotype and fate of primed T cells.
Immunology. 123: 480-90.
Canalli, A.A. et al. (2011) Participation of Mac-1, LFA-1 and VLA-4 integrins in the in vitro adhesion of sickle cell disease neutrophils to endothelial layers, and reversal of adhesion by simvastatin.
Haematologica. 96: 526-33.
Foster, G.R. et al. (2004) IFN-alpha subtypes differentially affect human T cell motility.
J Immunol. 173: 1663-70.
Haworth, O. et al. (2008) A role for the integrin alpha6beta1 in the differential distribution of CD4 and CD8 T-cell subsets within the rheumatoid synovium.
Rheumatology (Oxford). 47: 1329-34.
Nussbaum, G. et al. (2006) Peptide p277 of HSP60 signals T cells: inhibition of inflammatory chemotaxis.
Int Immunol. 18: 1413-9.
Peled, A. et al. (2002) Immature leukemic CD34+CXCR4+ cells from CML patients have lower integrin-dependent migration and adhesion in response to the chemokine SDF-1.
Stem Cells. 20: 259-66.
Ross, E.A. et al. (2006) Interaction between integrin alpha9beta1 and vascular cell adhesion molecule-1 (VCAM-1) inhibits neutrophil apoptosis.
Blood. 107: 1178-83.
Rosseau, S. et al. (2005) Moraxella catarrhalis--infected alveolar epithelium induced monocyte recruitment and oxidative burst.
Am J Respir Cell Mol Biol. 32: 157-66.
Spring, F.A.et al. (2001) Intercellular adhesion molecule-4 binds alpha(4)beta(1) and alpha(V)-family integrins through novel integrin-binding mechanisms.
Blood. 98: 458-66.
Hyduk, S.J. et al. (2011) Talin-1 and kindlin-3 regulate alpha4beta1 integrin-mediated adhesion stabilization, but not G protein-coupled receptor-induced affinity upregulation.
J Immunol. 187: 4360-8.
Caldwell, J.M. et al. (2017) Cadherin 26 is an alpha integrin-binding epithelial receptor regulated during allergic inflammation.
Mucosal Immunol. Jan 4. [Epub ahead of print]
Uitterdijk, A. et al. (2017) Time course of VCAM-1 expression in reperfused myocardial infarction in swine and its relation to retention of intracoronary administered bone marrow-derived mononuclear cells.
PLoS One. 12 (6): e0178779.