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: 1*PBS (pH7.4), 1% BSA, 40% Glycerol, 0.2% Proclean 950.
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: Application: Immunofluorescence (IHC-Fr)

Species: Mouse
Tissue: Cerebellum
Sample: Frozen section

Antigen retrieval: Recommend. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven.

Wash buffer: 1× TBST
Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.

Primary antibody: HA723738, 1/5,000, overnight at 4℃.
Secondary antibody: Goat anti-Rabbit IgG (iFluor™ 488), HA1121, 1/1,000, 1.5 hours at room temperature.
HA1121_2.jpg Fig2: Application: Immunofluorescence (IHC-Fr)

Species: Rat
Tissue: Cerebellum
Sample: Frozen section

Antigen retrieval: Recommend. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven.

Wash buffer: 1× TBST
Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.

Primary antibody: HA723738, 1/5,000, overnight at 4℃.
Secondary antibody: Goat anti-Rabbit IgG (iFluor™ 488), HA1121, 1/1,000, 1.5 hours at room temperature.
HA1121_3.jpg Fig3: Application: Immunofluorescence (IF-tissue)

Species: Rat
Tissue: Cerebellum
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, sodium citrate buffer (pH 6.0), 2 minutes at high pressure.

Wash buffer: 1× TBST
Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.
Primary antibody: Antibody A: 1/1,000 (NeuN , ET1602-12, rabbit, Green), Antibody B: 1/1,000 (GFAP, EM140707, mouse, Red), overnight at 4℃.
Secondary antibody: Goat anti-Rabbit IgG (iFluor™ 488), HA1121, 1/1,000; Goat anti-Mouse IgG (iFluor™ 594), HA1126, 1/1,000, 1.5 hours at room temperature.
HA1121_4.jpg Fig4: Application: Immunocytochemistry (IF-cell)

Species: Human
Sample: HeLa (Human cervix adenocarcinoma epithelial cell)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.

Primary antibody: beta Actin (HA722023), 1/250, overnight at 4℃.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), , 1/1,000, 45 minutes at room temperature.
Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).
HA1121_5.jpg Fig5: Application: Immunocytochemistry (IF-cell)

Species: Human
Sample: NIH/3T3 (Mouse fibroblast)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.

Primary antibody: beta Actin (HA722023), 1/100, overnight at 4℃.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1/1,000, 45 minutes at room temperature.
Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).
HA1121_6.jpg Fig6: Application: Immunocytochemistry (IF-cell)

Species: Human
Sample: L6 (Rat skeletal muscle myoblast)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.

Primary antibody: beta Actin (HA722023), 1/100, overnight at 4℃.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), , 1/1,000, 45 minutes at room temperature.
Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).
HA1121_7.jpg Fig7: Application: Flow Cytometry

Species: Human
Sample: MOLT-4 (Human acute T-Cell leukemia cell)

Blocking: 1% BSA + 10% normal goat serum + human FcR (Miltenyi Biotec), 15 minutes at room temperature.
Antibody dilution buffer: 1x PBS.

Primary antibody: CD3 (HA724302, 1 μg/mL, Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488), HA1121, 1/1,000, 15 minutes at room temperature.
HA1121_8.jpg Fig8: Application: Flow Cytometry (Intra)

Species: Human
Sample: HeLa (Human cervix adenocarcinoma epithelial cell)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Tween-20, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature.
Antibody dilution buffer: 1x PBS.

Primary antibody: beta actin (HA722023, 1 μg/mL, Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1/1,000, 15 minutes at room temperature.
HA1121_9.jpg Fig9: Western blot analysis of Goat Anti-Rabbit IgG - iFluor™ 488 Secondary Antibody (HA1121) at 1/5,000 dilution.

Lane 1: MCF7 (Human breast cancer cell) cell lysate (20 µg/Lane)
Lane 2: Jurkat (Human T-lymphoblastic cells) 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; ECL: K1801

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: Hsp90 beta (ET1605-56), 1/20,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature
Secondary antibody: Goat anti-Rabbit IgG (iFluor™ 488), HA1121, 1/5,000 in 5% NFDM/TBST, 1 hour at room temperature
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.