CD68 Recombinant Rabbit Monoclonal Antibody [PSH05-47]
cat.: HA722285
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Mouse, Rat
Applications: WB, IF-Cell, IHC-P, IHC-Fr, IF-Tissue
Clonality: Monoclonal
Clone number: PSH05-47
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 35 kDa
Isotype: IgG
Immunogen: Recombinant protein within Mouse CD68 aa 21-326.
Positive control: Mouse spleen tissue, rat spleen tissue, RAW264.7 cell lysate, RAW264.7, M‑NFS‑60 cell lysate, J774A.1 cell lysate, Rat spleen tissue lysate.
Subcellular location: Cell membrane; Endosome membrane, Lysosome membrane.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  IHC-Fr
  IF-Tissue
1:1,000-1:5,000
1:100
1:2,000
1:500
1:200
Uniprot #: SwissProt: P31996 Mouse
Entrez Gene: 287435 Rat
Alternative names: CD 68 CD68 CD68 antigen CD68 molecule CD68_HUMAN DKFZp686M18236 gp11 Gp110 LAMP4 Macrophage antigen CD68 (microsialin) MACROPHAGE ANTIGEN CD68 Macrosialin SCARD1 Scavenger receptor class D member 1
Images
HA722285_1.jpg Fig1: Immunofluorescence analysis of frozen mouse spleen tissue with Rabbit anti-CD68 antibody (HA722285) 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 (HA722285, 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).
HA722285_2.jpg Fig2: Immunofluorescence analysis of frozen mouse liver tissue with Rabbit anti-CD68 antibody (HA722285) 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 (HA722285, 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).
HA722285_3.jpg Fig3: Immunofluorescence analysis of frozen rat liver tissue with Rabbit anti-CD68 antibody (HA722285) 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 (HA722285, 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).
HA722285_4.jpg Fig4: 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.
HA722285_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded rat 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.
HA722285_6.jpg Fig6: 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: 35 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.
HA722285_7.jpg Fig7: Western blot analysis of CD68 on different lysates with Rabbit anti-CD68 antibody (HA722285) at 1/5,000 dilution.

Lane 1: RAW264.7 cell lysate (20 µg/Lane)
Lane 2: M‑NFS‑60 cell lysate (20 µg/Lane)
Lane 3: J774A.1 cell lysate (20 µg/Lane)
Lane 4: NIH/3T3 cell lysate (negative) (20 µg/Lane)
Lane 5: Neuro-2a cell lysate (negative) (20 µg/Lane)
Lane 6: Rat spleen tissue lysate (no heat) (40 µg/Lane)

Predicted band size: 35 kDa
Observed band size: 90-200 kDa

Exposure time: Lane 1-5: 12 seconds; Lane 6: 40 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 1/5,000 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.
HA722285_8.jpg Fig8: Immunofluorescence analysis of paraffin-embedded mouse spleen tissue labeling CD68 with Rabbit anti-CD68 antibody (HA722285) at 1/200 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 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).
HA722285_9.jpg Fig9: 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.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.