Anti-SIGIRR antibody [EPR22438-93]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIGIRR antibody [EPR22438-93] (AB228977)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling SIGIRR with ab228977 at 1/1000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human colon (PMID : 21077278) The section was incubated with ab228977 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIGIRR antibody [EPR22438-93] (AB228977)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling SIGIRR with ab228977 at 1/1000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human kidney (PMID : 14715412, 12925853) The section was incubated with ab228977 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIGIRR antibody [EPR22438-93] (AB228977)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling SIGIRR with ab228977 at 1/1000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse kidney (PMID : 14715412, 12925853). The section was incubated with ab228977 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIGIRR antibody [EPR22438-93] (AB228977)

Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling SIGIRR with ab228977 at 1/1000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat kidney (PMID : 14715412, 12925853). The section was incubated with ab228977 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-SIGIRR antibody [EPR22438-93] (AB228977)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized MEF (mouse embryonic fibroblast (immortalized)) (Left) / Hepa1-6 (mouse hepatoma epithelial cell line) (Right) cells labelling SIGIRR with ab228977 at 1/600 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat anti rabbit IgG (Alexa Fluor^� 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

Negative control : MEF (PMID 12925853).

• IP

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Immunoprecipitation - Anti-SIGIRR antibody [EPR22438-93] (AB228977)

IGFBP5 was immunoprecipitated from 0.35 mg Hepa1-6 (mouse hepatoma epithelial cell line) whole cell lysate 10μg with ab254324 at 1/30 dilution. Western blot was performed on the immunoprecipitate using ab228977 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used as the secondary antibody at 1/5000 dilution.

Lane 1 : Hepa1-6 (mouse hepatoma epithelial cell) whole cell lysate 10μg

Lane 2 : ab254324 IP in Hepa1-6 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab228977 in Hepa1-6 whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 5 seconds.

All lanes:

Immunoprecipitation - Anti-SIGIRR antibody [EPR22438-93] (ab228977)

Predicted band size: 46 kDa

Observed band size: 80 kDa

false

• WB

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Western blot - Anti-SIGIRR antibody [EPR22438-93] (AB228977)

The different apparent molecular weights are due to different levels of glycosylation. (PMID : 26344057).

Exposure times.
Lane 1 : 48 seconds; Lane 2 : 7.75 seconds; Lane 3 : 70 seconds; Lane 4 : 3 minutes.

Blocking/diluting buffer and concentration : 5% NFDM/TBST.

All lanes:

Western blot - Anti-SIGIRR antibody [EPR22438-93] (ab228977) at 1/1000 dilution

Lane 1:

Human fetal kidney tissue lysate at 20 µg

Lane 2:

Rat liver tissue lysate at 20 µg

Lane 3:

Mouse kidney tissue lysate at 20 µg

Lane 4:

Mouse spleen tissue lysate at 20 µg

Secondary

Lane 1:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Lanes 2 - 4:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 46 kDa

Observed band size: 80 kDa

false

• WB

Unknown

Western blot - Anti-SIGIRR antibody [EPR22438-93] (AB228977)

The different apparent molecular weights are due to different levels of glycosylation. (PMID : 26344057).

Negative control : MEF (PMID 12925853)

Blocking/Diluting buffer and concentration : 5% NFDM/TBST.

Exposure times.

Lane 1 : 15 seconds; Lanes 2-6 : 3 minutes.

All lanes:

Western blot - Anti-SIGIRR antibody [EPR22438-93] (ab228977) at 1/1000 dilution

Lanes 1 and 5:

Hepa1-6 (mouse hepatoma epithelial cell), whole cell lysate at 20 µg

Lane 2:

MEF (immortalized mouse embryonic fibroblasts), whole cell lysate at 20 µg

Lane 3:

Mouse stomach tissue lysate at 20 µg

Lane 4:

Mouse pancreas tissue lysate at 20 µg

Lane 6:

Rat stomach tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 46 kDa

Observed band size: 80 kDa

false

• WB

Unknown

Western blot - Anti-SIGIRR antibody [EPR22438-93] (AB228977)

The molecular weight observed is consistent with what has been described in the literature (PMID : 26344057).

Blocking/Diluting buffer and concentration : 5% NFDM/TBST

Exposure time : 92 seconds.

All lanes:

Western blot - Anti-SIGIRR antibody [EPR22438-93] (ab228977) at 1/1000 dilution

Lane 1:

Mouse kidney tissue lysate at 20 µg

Lane 2:

Mouse kidney treated lysate treated with with PNGase F at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 46 kDa

Observed band size: 46 kDa,80 kDa

false