Anti-Integrin beta 1 antibody [RM1259]

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Integrin beta 1 antibody [RM1259] (AB322405)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HepG2(human hepatocellular carcinoma epithelial cell) cells labelling Integrin beta 1 with ab322405 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin beta 1 antibody [RM1259] (AB322405)

Immunohistochemical analysis of paraffin-embedded Human hepatocellular carcinoma tissue labeling Integrin beta 1 with ab322405 at 1/2000 (0.249 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human hepatocellular carcinoma in (A) ab322405 and (B) ab183666. The image on (A) ab322405 and (B) ab183666 are treated with Anti-ITGB1 antibody at 1/2000 dilution. The section was incubated with ab322405 or ab183666 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Integrin beta 1 antibody [RM1259] (AB322405)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Parental HeLa(human cervical adenocarcinoma epithelial cell) (Green)/ITGB1 KO (human Integrin beta-1 knock out) HeLa (Magenta) cells labelling Integrin beta 1 with ab322405 at 1/50 dilution (1ug) / Magenta and Green compared with a Rabbit monoclonal IgG (ab172730) (Black) and Grey isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin beta 1 antibody [RM1259] (AB322405)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Integrin beta 1 with ab322405 at 1/2000 (0.249 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human colon in (A) ab322405 and (B) ab183666.
The image on (A) ab322405 and (B) ab183666 are treated with Anti-ITGB1 antibody at 1/2000 dilution. The section was incubated with ab322405 or ab183666 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Integrin beta 1 antibody [RM1259] (AB322405)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized ITGB1 KO HeLa (ITGB1 knockout human cervical adenocarcinoma epithelial cell), cells labelling Integrin beta 1 with ab322405 at 1/1000 (0.497 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green).

Confocal image showing cytoplasmic staining in wildtype HeLa cells, and showing no staining in ITGB1 knockout HeLa cells (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/ml dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Integrin beta 1 antibody [RM1259] (AB322405)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) cells labelling Integrin beta 1 with ab322405 at 1/1000 (0.497 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody ab150081 at 1/1000 2ug/ml dilution (Green).

Confocal image showing cytoplasmic and membranous staining in HepG2 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin ab195889 at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/ml dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-Integrin beta 1 antibody [RM1259] (AB322405)

Integrin beta 1 was immunoprecipitated from 0.35 mg HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate with ab322405 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab322405 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate
Lane 2 : ab322405 IP in HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab322405 in HepG2 whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-Integrin beta 1 antibody [RM1259] (ab322405) at 1/30 dilution

All lanes:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 32s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin beta 1 antibody [RM1259] (AB322405)

Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling Integrin beta 1 with ab322405 at 1/2000 (0.249 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat kidney. The section was incubated with ab322405 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Integrin beta 1 antibody [RM1259] (AB322405)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized PC-12(rat adrenal gland pheochromocytoma cell) cells labelling Integrin beta 1 with ab322405 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Integrin beta 1 antibody [RM1259] (AB322405)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Raw264.7(mouse Abelson murine leukemia virus-induced tumor macrophage) cells labelling Integrin beta 1 with ab322405 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin beta 1 antibody [RM1259] (AB322405)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling Integrin beta 1 with ab322405 at 1/2000 (0.249 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse kidney. The section was incubated with ab322405 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IP

Supplier Data

Immunoprecipitation - Anti-Integrin beta 1 antibody [RM1259] (AB322405)

Integrin beta 1 was immunoprecipitated from 0.35 mg PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate with ab322405 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab322405 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate
Lane 2 : ab322405 IP in PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab322405 in PC-12 whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-Integrin beta 1 antibody [RM1259] (ab322405) at 1/30 dilution

All lanes:

PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 15s

• IP

Supplier Data

Immunoprecipitation - Anti-Integrin beta 1 antibody [RM1259] (AB322405)

Integrin beta 1 was immunoprecipitated from 0.35 mg RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate with ab322405 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab322405 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 2 : ab322405 IP in RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab322405 in RAW 264.7 whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-Integrin beta 1 antibody [RM1259] (ab322405) at 1/30 dilution

All lanes:

RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 24s

• WB

Supplier Data

Western blot - Anti-Integrin beta 1 antibody [RM1259] (AB322405)

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

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 34646995; PMID : 19940135 ).

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

In Western blot, ab322405 was shown to bind specifically to ITGB1. A band was observed at 110-140 kDa in wild-type HeLa cell lysates with whereas no signal observed at this size in ITGB1 knockout cell line.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Left panel : Lanes 1-6 : 81 seconds; Lanes 7-8 : 37 seconds Right panel : 26 seconds

Observed MW(kDa) 110-140.

All lanes:

Western blot - Anti-Integrin beta 1 antibody [RM1259] (ab322405) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

ITGB1 knockout HeLa whole cell lysate at 20 µg

Lane 3:

U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 5:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 6:

IMR-32 (human neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 7:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 8:

C2C12 (mouse myoblast) whole cell 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

Observed band size: 110-140 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-Integrin beta 1 antibody [RM1259] (AB322405)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Observed MW(kDa) 110-140.

All lanes:

Western blot - Anti-Integrin beta 1 antibody [RM1259] (ab322405) at 1/1000 dilution

Lane 1:

Human liver tissue lysate at 20 µg

Lane 2:

Mouse heart tissue lysate at 20 µg

Lane 3:

Mouse kidney tissue lysate at 20 µg

Lane 4:

Mouse spleen tissue lysate at 20 µg

Lane 5:

Mouse liver tissue lysate at 20 µg

Lane 6:

Rat heart tissue lysate at 20 µg

Lane 7:

Rat kidney tissue lysate at 20 µg

Lane 8:

Rat spleen tissue lysate at 20 µg

Lane 9:

Rat liver 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

Observed band size: 110-140 kDa,36 kDa

false

Exposure time: 81s

• WB

Supplier Data

Western blot - Anti-Integrin beta 1 antibody [RM1259] (AB322405)

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

Observed MW(kDa) 110-140.

All lanes:

Western blot - Anti-Integrin beta 1 antibody [RM1259] (ab322405) at 1/1000 dilution

Lane 1:

Human cerebellum tissue lysate at 20 µg

Lane 2:

Human skeletal muscle tissue lysate at 20 µg

Lane 3:

Human spleen 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

Observed band size: 110-140 kDa

false

Exposure time: 26s

• WB

Supplier Data

Western blot - Anti-Integrin beta 1 antibody [RM1259] (AB322405)

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

Exposure times : Left panel : 26 seconds; right panel : 15 seconds.

Observed molecular weight 110-140 kDa.

All lanes:

Western blot - Anti-Integrin beta 1 antibody [RM1259] (ab322405) at 1/1000 dilution

Lane 1:

C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

Lane 2:

RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 3:

PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate at 20 µg

Lane 4:

NIH/3T3 (mouse embryonic fibroblast) whole cell 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

Observed band size: 110-140 kDa

false