iFluor™ 488 Conjugated Goat anti-rabbit IgG polyclonal Antibody
cat.: HA1121
Product Type: Goat polyclonal IgG, secondary antibodies
Species reactivity: Rabbit
Applications: IF-Cell, IF-Tissue, FC, IHC-Fr, WB
Clonality: Polyclonal
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: Preservative: 0.02% Sodium azide Constituents: 30% Glycerol, 1% BSA, 68.98% PBS
Concentration: 2ug/ul
Purification: Immunogen affinity purified.
Isotype: IgG
Immunogen: Rabbit IgG (H+L).
Recommended Dilutions:
  IF-Cell
  IF-Tissue
  FC
  IHC-Fr
  WB
1:500-1:1,000
1:500-1:1,000
1:500-1:1,000
1:500-1:1,000
1:5,000
Images
HA1121_1.jpg Fig1: Immunofluorescence analysis of paraffin-embedded rat cerebellum tissue labeling NeuN (ET1602-12) and GFAP (EM140707).
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 antibodies NeuN (ET1602-12, green) at 1/50 dilution and GFAP (EM140707, red) at 1/500 dilution at +4℃ overnight, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) and Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) were used as the secondary antibodies at 1/1,000 dilution. DAPI was used as nuclear counterstain.
HA1121_2.jpg Fig2: Immunocytochemistry analysis of HeLa cells labeling beta Actin with Rabbit anti-beta Actin antibody (HA722023) 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-beta Actin antibody (HA722023) 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.
HA1121_3.jpg Fig3: Immunocytochemistry analysis of NIH/3T3 cells labeling beta Actin with Rabbit anti-beta Actin antibody (HA722023) 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-beta Actin antibody (HA722023) 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.
HA1121_4.jpg Fig4: Immunocytochemistry analysis of L6 cells labeling beta Actin with Rabbit anti-beta Actin antibody (HA722023) 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-beta Actin antibody (HA722023) 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.
HA1121_5.jpg Fig5: Flow cytometric analysis of HeLa cells labeling beta Actin.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA722023, 1μg/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).
HA1121_6.jpg Fig6: Flow cytometric analysis of NIH/3T3 cells labeling beta Actin.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA722023, 1μg/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).
HA1121_7.jpg Fig7: Flow cytometric analysis of C6 cells labeling beta Actin.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA722023, 1μg/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).
HA1121_8.jpg Fig8: Western blot analysis of Goat Anti-Rabbit IgG - iFluor™ 488 Secondary Antibody (HA1121) at 1/5,000 dilution.

Lane 1: MCF7 cell lysate (20 µg/Lane)
Lane 2: Jurkat cell lysate (20 µg/Lane)
Lane 3: Mouse testis tissue lysate (40 µg/Lane)
Lane 4: Rat heart tissue lysate (40 µg/Lane)

Exposure time: 2 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 (ET1605-56) at 1/20,000 dilution was used at room temperature for 2 hours. Goat Anti-Rabbit IgG - iFluor™ 488 Secondary Antibody (HA1121) at 1/5,000 dilution was used for 1 hour at room temperature.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.