Anti-GOLM1/GOLPH2 antibody [EPR29022-61] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GOLM1/GOLPH2 antibody [EPR29022-61] - BSA and Azide free (AB320738)

This data was developed using ab320737, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labeling GOLM1/GOLPH2 with ab320737 at 1/2000 (0.246 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Low expression tissue : Nearly no staining in human skeletal muscle. The section was incubated with ab320737 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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GOLM1/GOLPH2 antibody [EPR29022-61] - BSA and Azide free (AB320738)

This data was developed using ab320737, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded (A) Wild-type HEK-293T (human embryonic kidney epithelial cell) cell pellets; (B) GOLM1 (GOLPH2) knockout HEK-293T cell pellets tissue labeling GOLM1/GOLPH2 with ab320737 at 1/2000 (0.246 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining in (A) Wild-type HEK-293T (human embryonic kidney epithelial cell) cell pellets, and no signal in (B) GOLM1 (GOLPH2) knockout HEK-293T cell pellets. The section was incubated with ab320737 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-GOLM1/GOLPH2 antibody [EPR29022-61] - BSA and Azide free (AB320738)

This data was developed using ab320737, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HEK293T (human embryonic kidney epithelial cell) cells labelling GOLM1/GOLPH2 with ab320737 at 1/500 (0.984 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing golgi staining in parental HEK293T cell line (shown in green) and no staining in GOLM1 (GOLPH2) knockout HEK-293T cell line. The counterstain was golgi marker which was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). ab169276 Anti-GM130 mouse polyclonal antibody - cis-Golgi Marker was used to counterstain tubulin at 1/200 (5 ug/ml) dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 (5 ug/ml) dilution (Magenta). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GOLM1/GOLPH2 antibody [EPR29022-61] - BSA and Azide free (AB320738)

This data was developed using ab320737, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human prostatic hyperplasia tissue labeling GOLM1/GOLPH2 with ab320737 at 1/2000 (0.246 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining in human prostatic hyperplasia. The section was incubated with ab320737 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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GOLM1/GOLPH2 antibody [EPR29022-61] - BSA and Azide free (AB320738)

This data was developed using ab320737, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human appendix tissue labeling GOLM1/GOLPH2 with ab320737 at 1/2000 (0.246 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining in human appendix. The section was incubated with ab320737 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

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Immunoprecipitation - Anti-GOLM1/GOLPH2 antibody [EPR29022-61] - BSA and Azide free (AB320738)

This data was developed using ab320737, the same antibody clone in a different buffer formulation. GOLM1/GOLPH2 was immunoprecipitated from 0.35 mg 293T (human embryonic kidney epithelial cell) whole cell lysate with ab320737 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab320737 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : 293T whole cell lysate Lane 2 : ab320737 IP in 293T whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab320737 in 293T whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 10 seconds.

All lanes:

Immunoprecipitation - Anti-GOLM1/GOLPH2 antibody [EPR29022-61] (<a href='/en-us/products/primary-antibodies/golm1-golph2-antibody-epr29022-61-ab320737'>ab320737</a>) at 1/30 dilution

All lanes:

293T (human embryonic kidney 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: 10s

• WB

Supplier Data

Western blot - Anti-GOLM1/GOLPH2 antibody [EPR29022-61] - BSA and Azide free (AB320738)

This data was developed using ab320737, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST. Negative control : HepG2. In Western blot, ab320737 was shown to bind specifically to GOLM1 (GOLPH2). Target of interest was observed at 75 kDa in wild-type HEK-293T cell lysates (lane 1) with no signal observed at this size in the GOLM1 (GOLPH2) knockout cell line (lane 2) (knockout cell line ab266505 / knockout cell lysate ab257976). The identity of the band above 75KDa in lanes 5 and 7 is unknown. In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control  (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-GOLM1/GOLPH2 antibody [EPR29022-61] (<a href='/en-us/products/primary-antibodies/golm1-golph2-antibody-epr29022-61-ab320737'>ab320737</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

Western blot - Human GOLM1 (GOLPH2) knockout HEK-293T cell lysate (<a href='/en-us/products/cell-lysates/human-golm1-golph2-knockout-hek-293t-cell-lysate-ab257976'>ab257976</a>) at 20 µg

Lane 3:

U-2 OS (human bone osteosarcoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

LNCaP (human prostate carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

Huh7 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 6:

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

Lane 7:

HeLa (human cervical adenocarcinoma epithelial cell) 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: 75 kDa,36 kDa

false

Exposure time: 70s

• WB

Supplier Data

Western blot - Anti-GOLM1/GOLPH2 antibody [EPR29022-61] - BSA and Azide free (AB320738)

This data was developed using ab320737, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST. Negative control : skeletal muscle In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control  (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-GOLM1/GOLPH2 antibody [EPR29022-61] (<a href='/en-us/products/primary-antibodies/golm1-golph2-antibody-epr29022-61-ab320737'>ab320737</a>) at 1/1000 dilution

Lane 1:

Human lung cancer tissue lysate at 20 µg

Lane 2:

Human prostate tissue lysate at 20 µg

Lane 3:

Human skeletal muscle tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 75 kDa,36 kDa

false

Exposure time: 48s