MA4400

antibody from Invitrogen Antibodies

Targeting: CD44 CD44R, CSPG8, HCELL, IN, MC56, MDU2, MDU3, MIC4, Pgp1

Provider product page for MA4400

Western blot

Immunocytochemistry

Immunoprecipitation

Immunohistochemistry

Flow cytometry

Blocking/Neutralizing

Other assay

Antibody data

Antibody Data
Antigen structure
References [27]
Comments [0]
Validations
Immunocytochemistry [5]
Flow cytometry [2]
Other assay [7]

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Product number: MA4400 - Provider product page
Provider: Invitrogen Antibodies
Product name: CD44 Monoclonal Antibody (Hermes-1)
Antibody type: Monoclonal
Antigen: Other
Description: MA4400 targets CD44 in flow cytometry, immunohistochemistry, immunoprecipitation, ICC/IF and neutralization applications and shows reactivity with bovine, goat, human, ovine, and rabbit samples. The MA4400 immunogen is 80 - 95 kDa lymphocyte surface glycoprotein H-CAM (CD44). This antibody is produced by injecting Rat IgG secreting hybridoma cells into the peritoneum of mice. The resulting ascites is collected from the mice and the antibody is purified. This product has been tested for endotoxins by limulus amoebocyte lysate (LAL) assay and contains an endotoxin concentration of less than or equal to 10 endotoxin units per milligram (EU/mg).
Reactivity: Human, Mouse, Bovine, Goat, Rabbit
Host: Rat
Isotype: IgG
Antibody clone number: Hermes-1
Vial size: 200 µg
Concentration: 1.0 mg/mL
Storage: -20°C

CD44 protein structure - MA4400 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of CD44 (green) showing staining in the membrane of Hela cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a CD44 monoclonal antibody (Product # MA4400) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4 ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of CD44 (green) showing staining in the membrane of MDA-MB-231 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a CD44 monoclonal antibody (Product # MA4400) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4 ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of CD44 (green) showing staining in the membrane of Hela cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a CD44 monoclonal antibody (Product # MA4400) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4 ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of CD44 (green) showing staining in the membrane of MDA-MB-231 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a CD44 monoclonal antibody (Product # MA4400) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4 ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of CD44 was performed using 70 % confluent log phase HeLa cells. The cells were fixed with 4 % paraformaldehyde for 10 minutes, permeabilized with 0.1 % Triton™ X-100 for 15 minutes, and blocked with 2 % BSA for 45 minutes at room temperature. The cells were labeled with CD44 Monoclonal Antibody (Hermes-1) (Product # MA4400, 1:100) in 0.1 % BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rat IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor™ Plus 488 (Product # A48269, 1:2,000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cell membrane like localization. Panel e represents no expression in MCF7. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 5 Gene expression of mast cell proteases by LUVA cells co-cultured (CC) with HLFs with or without RSV infection for 48 h (RSV CC and CTL CC, respectively). (A) LUVA cells co-cultured with RSV-infected HLFs demonstrated an increased expression of chymase mRNA compared to LUVA cells alone ( ** P = 0.01) and LUVA cells co-cultured with uninfected HLFs ( *** P = 0.002). RSV infection of LUVA cells alone did lead to a small increase in chymase expression at baseline ( * 1.7-fold, P = 0.03). Treatment with either 4-MU or neutralizing antibody to CD44 significantly decreased chymase expression by LUVA cells co-cultured with RSV-infected HLFs ( # P = 0.02; ## P = 0.01, respectively). (B) Expression of tryptase by LUVA cells co-cultured with RSV-infected HLFs was increased compared to LUVA cells alone ( ** P = 0.01) and LUVA cells co-cultured with uninfected HLFs ( *** P = 0.002). RSV infection of LUVA cells alone led to a slight increase in tryptase expression at baseline ( * 1.7-fold, P = 0.03). Treatment with either 4-MU or neutralizing antibody to CD44 decreased the expression of tryptase by LUVA cells co-cultured with RSV-infected HLFs ( # P = 0.02; ## P = 0.01, respectively). (C) Tryptase protein assayed by western blot confirmed a similar pattern as the gene expression analysis. Gene expression was normalized to GAPDH and is shown as normalized mean +- SD relative to control LUVA cells for N = 3-6 replicates/group.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 7 Rabbit meniscus healing in vivo by application of fibrin-loaded with CTGF and TGFbeta3 uS. By 1 wk post-op, the recruitment of CD44 + /CD90 + cells into the surgically created meniscus tears was observed with delivery of CTGF and TGFbeta3 uS in contrast to control ( a ) (arrow heads indicate remaining PLGA uS). The recruited CD44 + /CD90 + cells undergo differentiation into proCOL-I + /proCOL-Iialpha + fibrochondrocyte-like cells by 3 wks ( b ) (HR: healing region). CTGF-loaded fibrin with TGFbeta3 uS (C + T uS) successfully enhanced avascular meniscus healing ( c , d ) (Yellow arrows indicate the tear sites), supported by PR ( e ) and Saf-O staining ( f ). There were no noticeable damage on the articular surfaces both with the control ( g ) and C + T uS groups ( h ).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 5 OPN in NMO CSF promotes macrophage motility through integrin alphavbeta3. (A) Expression levels of integrin alphavbeta3 and CD44 on the surface of THP-1 cells were examined using FACS analysis. Prior to analysis, the cells were incubated with either the indicated Abs or control IgG, followed by incubation with Alexa Fluor-conjugated secondary Abs, as described in Materials and Methods. Inhibitory effect of 20 mug/ml integrin alphavbeta3 mAb (clone LM609, B), 100 muM GRGDSP peptide (RGD, C), and 20 mug/ml CD44 mAb (clone Hermes-1, D) on NMO or MS CSF-induced cell motility. The relative number of migrated cells to the lower surface of the membrane in the presence of 20 mug/ml control IgG (IgG, B and D) or 100 muM control GRGESP peptide (RGE, C) was taken as 100%. We used a two-tailed Student's t-test to assess differences. Data are means +- SDs from two or three independent experiments with triplicate data points per experiment. Other experimental conditions are described in the Materials and methods section.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. S1 Inhibitory effect of anti-CD44 mAb on MDA-MB231 cell motility. Two hundred microliters of 1 x 10 5 cells/ml human breast cancer cell line MDA-MB231 cells (2 x 10 4 cells in serum-free DMEM) were incubated with 10 mug/ml anti-CD44 mAb (clone Hermes-1) or control IgG. After 20 min incubation at room temperature, the cells were inoculated into each upper well of a chamber, and 750 mul of 10% serum containing DMEM medium was added to the lower well of a chamber. After 18 h incubation, the fixed and stained cells on the lower side of a membrane were photographed using a phase contrast microscope, and the numbers of migrated cells were counted. The relative number of migrated cells to the lower surface of the membrane in the presence of control IgG was taken as 100%. Data are represented as the mean +- SD from three independent experiments.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 2 Decidual-uNK cell co-cultures. A, Heatmap showing relative expression of genes coding putative HA receptors in different endometrial cell types in vivo. EC, endothelial cells; EnEC, endometrial epithelial cells. B-D, Flow cytometric analysis. B, Viability of uNK cells following MACS isolation was determined by FV660 exclusion. C, Purity of uNK cells after MACS isolation was determined by gating cells for CD56 and CD45. D, Histogram depicting CD56 + /CD44 + uNK cells after MACS isolation ( filled histogram ) vs negative control ( open histogram ). Representative dot plots or histogram are shown together with bar graphs (mean +- SEM) showing individual data points of three biological repeat experiments. E, Cytospin preparations of isolated uNK cells stained for DAPI (nucleus), CD56 and CD44. Co-localization of CD56 and CD44 is shown in bottom right panel. Scale = 100 uM. F, Schematic representation of the decidual-uNK cell co-culture killing assay. G-I, Analysis of SA-beta-Gal activity (G), senescent decidual cell markers (H), and decidual markers (I) in decidualizing EnSC co-cultured with or without uNK cells. The data show relative change to the indicated measurements in three biological repeat experiments compared to cultures decidualized with C+M for 8 days in the absence of uNK cells. Different letters above the bar graphs (mean +- SEM) indicate statistical differences ( P < .05) between groups (one-way ANOVA and Dunnett's multiple comparison test)

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 5 Gene expression of mast cell proteases by LUVA cells co-cultured (CC) with HLFs with or without RSV infection for 48 h (RSV CC and CTL CC, respectively). (A) LUVA cells co-cultured with RSV-infected HLFs demonstrated an increased expression of chymase mRNA compared to LUVA cells alone ( ** P = 0.01) and LUVA cells co-cultured with uninfected HLFs ( *** P = 0.002). RSV infection of LUVA cells alone did lead to a small increase in chymase expression at baseline ( * 1.7-fold, P = 0.03). Treatment with either 4-MU or neutralizing antibody to CD44 significantly decreased chymase expression by LUVA cells co-cultured with RSV-infected HLFs ( # P = 0.02; ## P = 0.01, respectively). (B) Expression of tryptase by LUVA cells co-cultured with RSV-infected HLFs was increased compared to LUVA cells alone ( ** P = 0.01) and LUVA cells co-cultured with uninfected HLFs ( *** P = 0.002). RSV infection of LUVA cells alone led to a slight increase in tryptase expression at baseline ( * 1.7-fold, P = 0.03). Treatment with either 4-MU or neutralizing antibody to CD44 decreased the expression of tryptase by LUVA cells co-cultured with RSV-infected HLFs ( # P = 0.02; ## P = 0.01, respectively). (C) Tryptase protein assayed by western blot confirmed a similar pattern as the gene expression analysis. Gene expression was normalized to GAPDH and is shown as normalized mean +- SD relative to control LUVA cells for N = 3-6 replicates/group.