Granulin Recombinant Rabbit Monoclonal Antibody [JE37-73]
cat.: HA721464
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: WB, IHC-P, IF-Cell, FC
Clonality: Monoclonal
Clone number: JE37-73
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 64 kDa
Isotype: IgG
Immunogen: Recombinant protein within Human Granulin aa 1-100 / 593.
Positive control: HepG2 cell lysate, HEK-293 cell lysate, TF-1 cell lysate, THP-1 cell lysate, human colon carcinoma tissue, human kidney tissue, THP-1.
Subcellular location: Secreted, Lysosome.
Recommended Dilutions:
  WB
  IHC-P
  IF-Cell
  FC

1:1,000
1:1,000
1:100
1:500-1:1,000
Uniprot #: SwissProt: P28799 Human
Alternative names: Acrogranin CLN11 GEP GP88 Granulin A Granulin B Granulin C Granulin D Granulin E Granulin epithelin Granulin F Granulin G Granulin-7 Granulins GRN GRN_HUMAN PC cell derived growth factor PCDGF PEPI PGRN Proepithelin Progranulin
Images
HA721464_1.jpg Fig1: Western blot analysis of Granulin on different lysates with Rabbit anti-Granulin antibody (HA721464) at 1/1,000 dilution.

Lane 1: HepG2 cell lysate
Lane 2: HEK-293 cell lysate
Lane 3: TF-1 cell lysate
Lane 4: THP-1 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 64 kDa
Observed band size: 50-75 kDa

Exposure time: 1 minutes; ECL: K1802;

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 (HA721464) 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.
HA721464_2.jpg Fig2: Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-Granulin antibody (HA721464) 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 (HA721464) 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.
HA721464_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Granulin antibody (HA721464) 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 (HA721464) 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.
HA721464_4.jpg Fig4: Immunocytochemistry analysis of THP-1 cells labeling Granulin with Rabbit anti-Granulin antibody (HA721464) at 1/100 dilution.

Cells were fixed in 100% methanol for 10 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and then blocked with 1% BSA for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Granulin antibody (HA721464) at 1/100 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. 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.
HA721464_5.jpg Fig5: Flow cytometric analysis of THP-1 cells labeling Granulin.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA721464, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.