WB
Western blot analysis of Recombinant Rabbit IgG on different lysates at 1/1,000 dilution. Lane 1: HeLa cell lysate Lane 2: Jurkat cell lysate Lane 3: 293T cell lysate Lane 4: NIH/3T3 cell lysate Lane 5: RAW264.7 cell lysate Lane 6: C6 cell lysate Lane 7: PC-12 cell lysate Lysates/proteins at 20 µg/Lane. Exposure time: 3 minutes; 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 at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human testis tissue with Recombinant Rabbit IgG at 1/200 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 at 1/200 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.ICC/IF
Immunocytochemistry analysis of NIH/3T3 cells labeling Recombinant Rabbit IgG 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 Recombinant Rabbit IgG at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.IP
Peroxiredoxin 5 was immunoprecipitated in 0.2mg HeLa cell lysate with Recombinant Rabbit IgG at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using Recombinant Rabbit IgG at 1/1,000 dilution. 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: Recombinant Rabbit IgG IP in HeLa cell lysate Lane 3: Recombinant Rabbit IgG Recombinant Rabbit monoclonal Antibody in HeLa cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 43 seconds.FC
Flow cytometric analysis of NIH/3T3 cells labeling Peroxiredoxin 5. Cells were fixed and permeabilized. Then stained with the primary antibody (1μg/mL) (green) compared with Recombinant Rabbit IgG (1μg/mL) (red). 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 at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).IHC
Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Recombinant Rabbit IgG at 1/200 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 at 1/200 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.IHC
Immunohistochemical analysis of paraffin-embedded rat testis tissue with Recombinant Rabbit IgG at 1/200 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 at 1/200 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.CHIP
Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells treated with 500ng/mL TSA for 4 hours with Histone H3 (acetyl K27) or Recombinant Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.| Product Name | Recombinant Rabbit IgG - Isotype control |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Small molecule. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | ChIPFCICC/IFIHCIPWB |
| WB:1:1000 IHC:1:200 ICC:1:100 IP:1-2μg/sample FC:1:1000 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Alternative Names | Ig gamma chain C region antibody Ig kappa-b4 chain C region antibody Ig kappa-b5 chain C region antibody Ig kappa-b9 chain C region antibody Ig lambda chain C region antibody K-BAS antibody |
|---|---|
| Function | IgG is a monomeric immunoglobulin, built of two heavy chains gamma and two light chains. Each molecule has two antigen binding sites. This is the most abundant immunoglobulin and is approximately equally distributed in blood and in tissue liquids, constituting 75% of serum immunoglobulins in humans. This is the only isotype that can pass through the placenta, thereby providing protection to the fetus in its first weeks of life before its own immune system has developed. It can bind to many kinds of pathogens, for example viruses, bacteria, and fungi, and protects the body against them by complement activation (classic pathway), opsonization for phagocytosis and neutralisation of their toxins. |
WB
Western blot analysis of Recombinant Rabbit IgG on different lysates at 1/1,000 dilution. Lane 1: HeLa cell lysate Lane 2: Jurkat cell lysate Lane 3: 293T cell lysate Lane 4: NIH/3T3 cell lysate Lane 5: RAW264.7 cell lysate Lane 6: C6 cell lysate Lane 7: PC-12 cell lysate Lysates/proteins at 20 µg/Lane. Exposure time: 3 minutes; 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 at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human testis tissue with Recombinant Rabbit IgG at 1/200 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 at 1/200 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.
ICC/IF
Immunocytochemistry analysis of NIH/3T3 cells labeling Recombinant Rabbit IgG 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 Recombinant Rabbit IgG at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.
IP
Peroxiredoxin 5 was immunoprecipitated in 0.2mg HeLa cell lysate with Recombinant Rabbit IgG at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using Recombinant Rabbit IgG at 1/1,000 dilution. 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: Recombinant Rabbit IgG IP in HeLa cell lysate Lane 3: Recombinant Rabbit IgG Recombinant Rabbit monoclonal Antibody in HeLa cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 43 seconds.
FC
Flow cytometric analysis of NIH/3T3 cells labeling Peroxiredoxin 5. Cells were fixed and permeabilized. Then stained with the primary antibody (1μg/mL) (green) compared with Recombinant Rabbit IgG (1μg/mL) (red). 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 at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
IHC
Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Recombinant Rabbit IgG at 1/200 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 at 1/200 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.
IHC
Immunohistochemical analysis of paraffin-embedded rat testis tissue with Recombinant Rabbit IgG at 1/200 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 at 1/200 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.
CHIP
Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells treated with 500ng/mL TSA for 4 hours with Histone H3 (acetyl K27) or Recombinant Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.| Application Notes | WB:1:1000 IHC:1:200 ICC:1:100 IP:1-2μg/sample FC:1:1000 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
