Product Type: | Antibody Sampler Kit |
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Species reactivity: | Mouse |
Applications: | WB |
Form: | Liquid |
Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
Storage buffer: | PBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
Uniprot #: | SwissProt: Q61549 Mouse | Q5Y4N8 Rat | P31996 Mouse | P42081 Human | P42082 Mouse | Q9QXH4 Mouse | P20702 Human | Q61830 Mouse | P05089 Human | Q61176 Mouse | P07824 Rat | P55008 Human | O70200 Mouse | P55009 Rat Entrez Gene: 287435 Rat | 56822 Rat | 291327 Rat |
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Fig1:
Western blot analysis of F4/80 on different lysates with Rabbit anti-F4/80 antibody (HA721520) at 1/1,000 dilution. Lane 1: RAW264.7 cell lysate (no heat) (20 µg/Lane) Lane 2: L929 cell lysate (no heat) (negative) (20 µg/Lane) Lane 3: Rat spleen tissue lysate (70℃ heat) (40 µg/Lane) Notice: no heat means the lysate is not boiled. Predicted band size: 102 kDa Observed band size: 160 kDa Exposure time: 2 minutes; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721520) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature. |
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Fig2:
Western blot analysis of CD68 on different lysates with Rabbit anti-CD68 antibody (HA722285) at 1/1,000 dilution. Lane 1: RAW264.7 cell lysate Lane 2: RAW264.7 cell lysate treated with deglycosylation Lysates/proteins at 20 µg/Lane. Predicted band size: 37 kDa Observed band size: 90/70 kDa Exposure time: 25 seconds; ECL: K1801; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722285) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig3:
Western blot analysis of CD86 on different lysates with Rabbit anti-CD86 antibody (ET1606-50) at 1/2,000 dilution. Lane 1: Daudi cell lysate, 15 µg/Lane Lane 2: Raji cell lysate, 15 µg/Lane Lane 3: Ramos cell lysate, 15 µg/Lane Lane 4: Jurkat cell lysate, 15 µg/Lane Lane 5: K-562 cell lysate, 15 µg/Lane Predicted band size: 38 kDa Observed band size: 70 kDa Exposure time: 2 minutes 37 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1606-50) at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature. |
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Fig4:
Western blot analysis of CD86 on different lysates with Rabbit anti-CD86 antibody (ET1606-50) at 1/1,000 dilution. Lane 1: Raji cell lysate (10 µg/Lane) Lane 2: Mouse lung tissue lysate (20 µg/Lane) Lane 3: Rat lung tissue lysate (20 µg/Lane) Predicted band size: 38 kDa Observed band size: 70 kDa Exposure time: 1 minute; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1606-50) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature. |
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Fig5:
Western blot analysis of CD11c on different lysates with Rabbit anti-CD11c antibody (HA722830) at 1/1,000 dilution. Lane 1: RAW264.7 cell lysate (20 µg/Lane) Lane 2:NIH/3T3 cell lysate (negative) (20 µg/Lane) Lane 3: C2C12 cell lysate (negative) (20 µg/Lane) Lane 4: Mouse spleen tissue lysate (30 µg/Lane) Lane 5: Mouse colon tissue lysate (30 µg/Lane) Predicted band size: 129 kDa Observed band size: 140 kDa Exposure time: Lane1-3: 3 minutes ; Lane 4-5: 25 seconds;ECL: K1801; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722830) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig6:
Western blot analysis of Mannose Receptor(CD206) on different lysates with Rabbit anti-Mannose Receptor(CD206) antibody (HA722892) at 1/2,000 dilution. Lane 1: RAW264.7 cell lysate Lane 2: RAW264.7 treated with 20ng/mL mIL-4 and 10ng/mL mIL-13 for 24 hours cell lysate (Macrophage M2 polarization, positive) Lane 3: RAW264.7 cell lysate Lane 4: RAW264.7 treated with 0.1μg/mL LPS for 6 hours cell lysate (Macrophage M1 polarization, negative) Lysates/proteins at 20 µg/Lane. Predicted band size: 165 kDa Observed band size: 200 kDa Exposure time: 59 seconds; ECL: K1801; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722892) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig7:
Western blot analysis of Mannose Receptor(CD206) on different lysates with Rabbit anti-Mannose Receptor(CD206) antibody (HA722892) at 1/1,000 dilution. Lane 1: Mouse lung tissue lysate (20 µg/Lane) Lane 2: Mouse spleen tissue lysate (20 µg/Lane) Lane 3: Rat brain tissue lysate (20 µg/Lane) Lane 4: Rat lung tissue lysate (20 µg/Lane) Lane 5: Rat liver tissue lysate (20 µg/Lane) Predicted band size: 165 kDa Observed band size: 200 kDa Exposure time: 42 seconds; ECL: K1801; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722892) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig8:
Western blot analysis of Iba1 on different lysates with Rabbit anti-Iba1 antibody (ET1705-78) at 1/5,000 dilution. Lane 1: THP-1 cell lysate, 20 µg/Lane Lane 2: HEK-293 cell lysate (negative), 20 µg/Lane Lane 3: Mouse spleen tissue lysate, 20 µg/Lane Lane 4: Rat spleen tissue lysate, 20 µg/Lane Predicted band size: 17 kDa Observed band size: 17 kDa Exposure time: 3 minutes; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1705-78) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:50,000 dilution was used for 1 hour at room temperature. |
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Fig9:
Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-F4/80 antibody (HA721520) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721520) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. |
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Fig10:
Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-CD68 antibody (HA722285) at 1/2,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722285) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. |
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Fig11:
Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-CD11c antibody (HA722830) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722830) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. |
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Fig12:
Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-Mannose Receptor(CD206) antibody (HA722892) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722892) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. |
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Fig13:
Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-Iba1 antibody (ET1705-78) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-78) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. |
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Fig14:
Immunocytochemistry analysis of RAW264.7 cells labeling CD68 with Rabbit anti-CD68 antibody (HA722285) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-CD68 antibody (HA722285) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig15:
Immunocytochemistry analysis of RAW264.7 cells treated with 20ng/mL mIL-4 and 10ng/mL mIL-13 for 24 hours (Macrophage M2 polarization, positive) labeling Mannose Receptor(CD206) with Rabbit anti-Mannose Receptor(CD206) antibody (HA722892) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Mannose Receptor(CD206) antibody (HA722892) at 1/50 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig16:
Immunocytochemistry analysis of RAW264.7 cells treated with 0.1μg/mL LPS for 6 hours (Macrophage M1 polarization, negative) labeling Mannose Receptor(CD206) with Rabbit anti-Mannose Receptor(CD206) antibody (HA722892) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Mannose Receptor(CD206) antibody (HA722892) at 1/50 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig17:
Immunocytochemistry analysis of RAW264.7 cells labeling Iba1 with Rabbit anti-Iba1 antibody (ET1705-78) at 1/250 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Iba1 antibody (ET1705-78) at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig18:
Immunofluorescence analysis of frozen mouse spleen tissue with Rabbit anti-F4/80 antibody (HA721520) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA721520, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue). |
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Fig19:
Immunofluorescence analysis of frozen mouse lymph nodes tissue with Rabbit anti-CD68 antibody (HA722285) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA722285, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue). |
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Fig20:
Immunofluorescence analysis of frozen mouse spleen tissue with Rabbit anti-CD11c antibody (HA722830) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA722830, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue). |
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Fig21:
Immunofluorescence analysis of frozen mouse spleen tissue with Rabbit anti-Mannose Receptor(CD206) antibody (HA722892) at 1/500 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA722892, green) at 1/500 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue). |
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Fig22:
Immunofluorescence analysis of frozen mouse brain tissue with Rabbit anti-Iba1 antibody (ET1705-78) at 1/500 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1705-78, green) at 1/500 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue). |