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SAGE1 Recombinant Rabbit Monoclonal Antibody [PSH05-89] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA751036

SAGE1 Recombinant Rabbit Monoclonal Antibody [PSH05-89] - BSA and Azide free

Application

  • WB

  • IHC-P

  • IF-Cell

  • IP

Reactivity

  • Human

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

SAGE1 Recombinant Rabbit Monoclonal Antibody [PSH05-89] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Human SAGE1 aa 75-733.

Species Reactivity

Human

Validated Applications

WB, IHC-P, IF-Cell, IP

Molecular Weight

Predicted band size: 99 kDa

Positive Control

K-562 cell lysates, K-562, human testis tissue.

Conjugation

unconjugated

Clone Number

PSH05-89

Product Features

Form

Liquid

Concentration

1mg/ml

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000

  • IHC-P

  • 1:1,000

  • IF-Cell

  • 1:500

  • IP

  • 1-2μg/sample

Target

Function

This gene belongs to a class of genes that are activated in tumors. These genes are expressed in tumors of different histologic types but not in normal tissues, except for spermatogenic cells and, for some, placenta. The proteins encoded by these genes appear to be strictly tumor specific, and hence may be excellent sources of antigens for cancer immunotherapy. This gene is expressed in sarcomas.

Background References

1. Miyahara Y., Naota H., Wang L., Hiasa A., Goto M., Watanabe M., Kitano S., Okumura S., Takemitsu T., Shiku H. Determination of cellularly processed HLA-A2402-restricted novel CTL epitopes derived from two cancer germ line genes, MAGE-A4 and SAGE. Clin Cancer Res 11:5581-5589 (2005)

2. Maheswaran E., Pedersen C.B., Ditzel H.J., Gjerstorff M.F. Lack of ADAM2, CALR3 and SAGE1 Cancer/Testis Antigen Expression in Lung and Breast Cancer. PLoS One 10:e0134967-e0134967 (2015)

Subcellular Location

Nuclear body. Nucleoplasm.

UNIPROT

SwissProt: Q9NXZ1 Human

Synonyms

Cancer/testis antigen 14 antibody

CT14 antibody

RP11-97N5.1 antibody

SAGE antibody

SAGE1 antibody

SAGE1_HUMAN antibody

Sarcoma antigen 1 antibody

Images

  • Western blot analysis of SAGE1 on K-562 cell lysates with Rabbit anti-SAGE1 antibody (<a href="/products/HA751036" style="font-weight: bold;text-decoration: underline;">HA751036</a>) at 1/1,000 dilution.<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 99 kDa<br />Observed band size: 120 kDa<br /><br />Exposure time: 20 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA751036" style="font-weight: bold;text-decoration: underline;">HA751036</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of SAGE1 on K-562 cell lysates with Rabbit anti-SAGE1 antibody (HA751036) at 1/1,000 dilution.

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 99 kDa
    Observed band size: 120 kDa

    Exposure time: 20 seconds; ECL: K1801;

    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 (HA751036) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of K-562 cells labeling SAGE1 with Rabbit anti-SAGE1 antibody (<a href="/products/HA751036" style="font-weight: bold;text-decoration: underline;">HA751036</a>) at 1/500 dilution.<br /><br />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 Rabbit anti-SAGE1 antibody (<a href="/products/HA751036" style="font-weight: bold;text-decoration: underline;">HA751036</a>) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) 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.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of K-562 cells labeling SAGE1 with Rabbit anti-SAGE1 antibody (HA751036) at 1/500 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 Rabbit anti-SAGE1 antibody (HA751036) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-SAGE1 antibody (<a href="/products/HA751036" style="font-weight: bold;text-decoration: underline;">HA751036</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA751036" style="font-weight: bold;text-decoration: underline;">HA751036</a>) at 1/1,000 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.

    Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-SAGE1 antibody (HA751036) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (HA751036) at 1/1,000 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.

  • SAGE1 was immunoprecipitated from 0.2 mg K-562 cell lysate with <a href="/products/HA751036" style="font-weight: bold;text-decoration: underline;">HA751036</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA751036" style="font-weight: bold;text-decoration: underline;">HA751036</a> at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: K-562 cell lysate (input)<br />Lane 2: <a href="/products/HA751036" style="font-weight: bold;text-decoration: underline;">HA751036</a> IP in K-562 cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/HA751036" style="font-weight: bold;text-decoration: underline;">HA751036</a> in K-562 cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 3 minutes; ECL: <a href="/products/K1801" style="font-weight: bold;text-decoration: underline;">K1801</a>

    SAGE1 was immunoprecipitated from 0.2 mg K-562 cell lysate with HA751036 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751036 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: K-562 cell lysate (input)
    Lane 2: HA751036 IP in K-562 cell lysate
    Lane 3: Rabbit IgG instead of HA751036 in K-562 cell lysate

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 3 minutes; ECL: K1801

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"