FAF1 Recombinant Rabbit Monoclonal Antibody [PSH22-51]
cat.: HA724301
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat, Monkey
Applications: WB, IF-Cell, IHC-P, FC, IP
Clonality: Monoclonal
Clone number: PSH22-51
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: 1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 74 kDa
Isotype: IgG
Immunogen: Recombinant protein within human FAF1 aa 1-530.
Positive control: THP-1 cell lysate, 293T cell lysate, HeLa cell lysate, U-2 OS cell lysate, COS-1 cell lysate, NIH/3T3 cell lysate, C6 cell lysate, Mouse brain tissue lysate, Mouse heart tissue lysate, Mouse kidney tissue lysate, Rat brain tissue lysate, Rat heart tissue lysate, Rat liver tissue lysate, HeLa, NIH/3T3, C6, human colon tissue, human ovarian carcinoma tissue, human testis tissue, mouse colon tissue, mouse testis tissue, rat colon tissue, rat testis tissue.
Subcellular location: Nucleus.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC
  IP
1:20,000
1:100
1:500-1:2,000
1:1,000
1-2μg/sample
Uniprot #: SwissProt: Q9UNN5 Human | P54731 Mouse | Q924K2 Rat
Alternative names: CGI 03 FAF1 FAF1 protein FAF1_HUMAN Fas (TNFRSF6) associated factor 1 FAS associated factor 1 Fas associated protein factor 1 FAS-associated factor 1 FLJ37524 hFAF1 HFAF1s Protein FAF1 TNFRSF6 associated factor 1 UBX domain protein 3A UBX domain-containing protein 12 UBX domain-containing protein 3A UBXD12 UBXN3A
Images
HA724301_1.jpg Fig1: Western blot analysis of FAF1 on different lysates with Rabbit anti-FAF1 antibody (HA724301) at 1/20,000 dilution.

Lane 1: THP-1 cell lysate (10 µg/Lane)
Lane 2: 293T cell lysate (10 µg/Lane)
Lane 3: HeLa cell lysate (10 µg/Lane)
Lane 4: U-2 OS cell lysate (10 µg/Lane)
Lane 5: COS-1 cell lysate (10 µg/Lane)
Lane 6: NIH/3T3 cell lysate (10 µg/Lane)
Lane 7: C6 cell lysate (10 µg/Lane)

Predicted band size: 74 kDa
Observed band size: 74 kDa

Exposure time: 4 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 (HA724301) at 1/20,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HA724301_2.jpg Fig2: Western blot analysis of FAF1 on different lysates with Rabbit anti-FAF1 antibody (HA724301) at 1/20,000 dilution.

Lane 1: Mouse brain tissue lysate (20 µg/Lane)
Lane 2: Mouse heart tissue lysate (20 µg/Lane)
Lane 3: Mouse kidney tissue lysate (20 µg/Lane)
Lane 4: Rat brain tissue lysate (20 µg/Lane)
Lane 5: Rat heart tissue lysate (20 µg/Lane)
Lane 6: Rat liver tissue lysate (20 µg/Lane)

Predicted band size: 74 kDa
Observed band size: 74 kDa

Exposure time: 10 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 (HA724301) at 1/20,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HA724301_3.jpg Fig3: Immunocytochemistry analysis of HeLa cells labeling FAF1 with Rabbit anti-FAF1 antibody (HA724301) 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-FAF1 antibody (HA724301) 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.
HA724301_4.jpg Fig4: Immunocytochemistry analysis of NIH/3T3 cells labeling FAF1 with Rabbit anti-FAF1 antibody (HA724301) 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-FAF1 antibody (HA724301) 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.
HA724301_5.jpg Fig5: Immunocytochemistry analysis of C6 cells labeling FAF1 with Rabbit anti-FAF1 antibody (HA724301) 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-FAF1 antibody (HA724301) 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.
HA724301_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-FAF1 antibody (HA724301) 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 (HA724301) 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.
HA724301_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma tissue with Rabbit anti-FAF1 antibody (HA724301) at 1/500 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 (HA724301) at 1/500 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.
HA724301_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-FAF1 antibody (HA724301) 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 (HA724301) 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.
HA724301_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-FAF1 antibody (HA724301) 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 (HA724301) 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.
HA724301_10.jpg Fig10: Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-FAF1 antibody (HA724301) 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 (HA724301) 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.
HA724301_11.jpg Fig11: Immunohistochemical analysis of paraffin-embedded rat colon tissue with Rabbit anti-FAF1 antibody (HA724301) 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 (HA724301) 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.
HA724301_12.jpg Fig12: Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-FAF1 antibody (HA724301) 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 (HA724301) 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.
HA724301_13.jpg Fig13: Flow cytometric analysis of HeLa cells labeling FAF1.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA724301, 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).
HA724301_14.jpg Fig14: Flow cytometric analysis of NIH/3T3 cells labeling FAF1.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA724301, 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).
HA724301_15.jpg Fig15: Flow cytometric analysis of C6 cells labeling FAF1.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA724301, 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).
HA724301_16.jpg Fig16: FAF1 was immunoprecipitated from 0.2 mg HeLa cell lysate with HA724301 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA724301 at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: HeLa cell lysate (input)
Lane 2: HA724301 IP in HeLa cell lysate
Lane 3: Rabbit IgG instead of HA724301 in HeLa cell lysate

Blocking/Dilution buffer: primary antibody dilution (K1803)
Exposure time: 6 seconds; ECL: K1801
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.