CARD9 Recombinant Rabbit Monoclonal Antibody [PSH0-41]
cat.: HA721343
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, FC
Clonality: Monoclonal
Clone number: PSH0-41
Form: Liquid
Storage condition: Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
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: 62.2 kDa
Isotype: IgG
Immunogen: Recombinant protein within human CARD9 aa 1-200 / 536.
Positive control: HL-60 cell lysate, THP-1 cell lysate, RAW264.7 cell lysate, PC-12 cell lysate, THP-1, RAW264.7, PC-12.
Subcellular location: Cytoplasm..
Recommended Dilutions:
  WB
  IF-Cell
  FC

1:1,000
1:100
1:500-1:1,000
Uniprot #: SwissProt: Q9H257 Human | A2AIV8 Mouse | A2AIV8 Rat
Alternative names: CANDF2 CARD9 CARD9_HUMAN Caspase recruitment domain family member 9 Caspase recruitment domain-containing protein 9 hCARD9
Images
HA721343_1.jpg Fig1: Western blot analysis of CARD9 on different lysates with Rabbit anti-CARD9 antibody (HA721343) at 1/1,000 dilution.

Lane 1: HL-60 cell lysate
Lane 2: THP-1 cell lysate
Lane 3: RAW264.7 cell lysate
Lane 4: PC-12 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 62.2 kDa
Observed band size: 70 kDa

Exposure time: 24 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 (HA721343) 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 1:200,000 dilution was used for 1 hour at room temperature.
HA721343_2.jpg Fig2: Immunocytochemistry analysis of THP-1 cells labeling CARD9 with Rabbit anti-CARD9 antibody (HA721343) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-CARD9 antibody (HA721343) 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.
HA721343_3.jpg Fig3: Immunocytochemistry analysis of RAW264.7 cells labeling CARD9 with Rabbit anti-CARD9 antibody (HA721343) at 1/100 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-CARD9 antibody (HA721343) 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 (HA601187, 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.
HA721343_4.jpg Fig4: Immunocytochemistry analysis of PC-12 cells labeling CARD9 with Rabbit anti-CARD9 antibody (HA721343) at 1/100 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-CARD9 antibody (HA721343) 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 (HA601187, 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.
HA721343_5.jpg Fig5: Flow cytometric analysis of THP-1 cells labeling CARD9.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA721343, 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).
HA721343_6.jpg Fig6: Flow cytometric analysis of RAW264.7 cells labeling CARD9.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA721343, 1/1,000) (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”.