Anti-PTP1B antibody [EPR22474] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTP1B antibody [EPR22474] - BSA and Azide free (AB245984)

Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling PTP1B with ab244207 at 1/1000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Higher cytoplasmic expression in human breast cancer than that of adjacent normal tissues (PMID : 27465552) is observed. Counterstained with hematoxylin.
Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab244207).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTP1B antibody [EPR22474] - BSA and Azide free (AB245984)

Immunohistochemical analysis of paraffin-embedded human colon cancer tissue labeling PTP1B with ab244207 at 1/1000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining in human colon cancer (PMID : 27752061) is observed. Counterstained with hematoxylin.
Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab244207).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-PTP1B antibody [EPR22474] - BSA and Azide free (AB245984)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Wild type and PTP1B-knockout HAP1 (Human chronic myelogenous leukemia near-haploid cell line) cells labeling PTP1B with ab244207 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing no staining in PTP1B-knockout HAP1 cells. The nuclear counterstain is DAPI (blue).
Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at a 1/200 dilution (red).
The negative control is the secondary antibody only.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab244207).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-PTP1B antibody [EPR22474] - BSA and Azide free (AB245984)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HCT 116 (human colorectal carcinoma epithelial cell) cell line labeling PTP1B with ab244207 at 1/50 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab244207).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-PTP1B antibody [EPR22474] - BSA and Azide free (AB245984)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HCT 116 (human colorectal carcinoma epithelial cell) cells labeling PTP1B with ab244207 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining in HCT 116 cells. The nuclear counterstain is DAPI (blue).
Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at a 1/200 dilution (red).
The negative control is the secondary antibody only.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab244207).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-PTP1B antibody [EPR22474] - BSA and Azide free (AB245984)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labeling PTP1B with ab244207 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining in HeLa cells. The nuclear counterstain is DAPI (blue).
Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at a 1/200 dilution (red).
The negative control is the secondary antibody only.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab244207).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-PTP1B antibody [EPR22474] - BSA and Azide free (AB245984)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cell line labeling PTP1B with ab244207 at 1/500 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab244207).

• IP

Unknown

Immunoprecipitation - Anti-PTP1B antibody [EPR22474] - BSA and Azide free (AB245984)

PTP1B was immunoprecipitated from 0.35 mg HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate with ab244207 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab244207 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.
Lane 1 : HCT 116 whole cell lysate 10 μg (Input).
Lane 2 : ab244207 IP in HCT 116 whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab244207 in HCT 116 whole cell lysate.
Blocking/Dilution buffer : 5% NFDM/TBST.
Exposure time : 30 seconds.

The observed MW is consistent with what described in the literatures. (PMID : 18253097; PMID : 11895943; PMID : 19797268).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab244207).

All lanes:

Immunoprecipitation - Anti-PTP1B antibody [EPR22474] (<a href='/en-us/products/primary-antibodies/ptp1b-antibody-epr22474-ab244207'>ab244207</a>)

Predicted band size: 50 kDa

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• WB

Lab

Western blot - Anti-PTP1B antibody [EPR22474] - BSA and Azide free (AB245984)

This data was developed using the same antibody clone in a different buffer formulation (ab244207).

Lanes 1- 2 : Merged signal (red and green). Green - ab244207 observed at 50 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab244207 was shown to react with PTP1B in wild-type HeLa cells in western blot. The band observed in CRISPR/Cas9 edited cell line ab265014 (CRISPR/Cas9 edited cell lysate ab257617) lane below 50kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and PTPN1 CRISPR/Cas9 edited HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab244207 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-PTP1B antibody [EPR22474] (<a href='/en-us/products/primary-antibodies/ptp1b-antibody-epr22474-ab244207'>ab244207</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

PTPN1 CRISPR/Cas9 edited HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human PTPN1 (PTP1B) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-ptpn1-ptp1b-knockout-hela-cell-line-ab265014'>ab265014</a>)

Predicted band size: 50 kDa

Observed band size: 50 kDa

false

• WB

Lab

Western blot - Anti-PTP1B antibody [EPR22474] - BSA and Azide free (AB245984)

This data was developed using the same antibody clone in a different buffer formulation (ab244207).

Lanes 1 - 4 : Merged signal (red and green). Green - ab244207 observed at 51 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

ab244207 was shown to react with PTP1B in wild-type HeLa cells in Western blot with loss of signal observed in PTPN1 knockout cell line ab265014 (PTPN1 knockout cell lysate 257617). Wild-type HeLa and PTPN1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5 % milk in TBS-T (0.1 % Tween^®) before incubation with ab244207 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-PTP1B antibody [EPR22474] (<a href='/en-us/products/primary-antibodies/ptp1b-antibody-epr22474-ab244207'>ab244207</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

PTPN1 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human PTPN1 (PTP1B) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-ptpn1-ptp1b-knockout-hela-cell-line-ab265014'>ab265014</a>)

Lane 3:

Jurkat cell lysate at 20 µg

Lane 4:

MCF7 cell lysate at 20 µg

Predicted band size: 50 kDa

Observed band size: 51 kDa

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