Name: Anti-SQSTM1 / p62 antibody [EPR18351] - BSA and Azide free
SKU: ab227992
Rating: 3 (1 reviews)

Anti-SQSTM1 / p62 [EPR18351] antibody (ab227992) is a rabbit monoclonal antibody that is provided in a PBS only formulation free from BSA and azide. Formulated to be conjugation-ready. It is used to detect SQSTM1 / p62 in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IP, IHC-P, ICC/IF. Suitable for Human samples.

- Specificity confirmed with SQSTM1 / p62 knockout cell line validation

-BSA,?sodium azide, and glycerol-free?for consistent conjugation with fluorochromes, enzymes and more

Images

Immunoprecipitation - Anti-SQSTM1 / p62 antibody [EPR18351] - BSA and Azide free (AB227992), expandable thumbnail

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: PBS
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	ICC/IF	IP	WB	Flow Cyt (Intra)	IHC-P
Human	Tested	Tested	Tested	Tested	Tested

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes ab199376 - Rabbit monoclonal IgG (Low endotoxin, Azide free), is suitable for use as an isotype control with this antibody.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Associated Products

Select an associated product type

15 products for Alternative Product

2 products for Alternative Version

Target data

See full target information SQSTM1

Function

The protein expressed by the SQSTM1 gene is an autophagy receptor essential for selective macroautophagy (aggrephagy). It acts as a bridge between polyubiquitinated cargo and autophagosomes, interacting with both the cargo and an autophagy modifier from the MAP1 LC3 family. Alongside WDFY3, it plays a role in forming and autophagically degrading cytoplasmic ubiquitin-containing inclusions and recruiting ubiquitinated proteins to nuclear PML bodies. SQSTM1 may regulate NFKB1 activation by TNF-alpha, nerve growth factor (NGF), and interleukin-1, and may influence titin/TTN signaling in muscle cells. It may also affect signaling cascades via ubiquitination and be involved in cell differentiation, apoptosis, immune response, and regulation of potassium channels. The protein is involved in endosome organization by retaining vesicles in the perinuclear cloud; ubiquitination by RNF26 allows it to attract vesicle-associated adapters, forming a molecular bridge to organize endosomal pathways. Additionally, it promotes the relocalization of Lys-63-linked ubiquitinated STING1 to autophagosomes and acts as an activator of the NFE2L2/NRF2 pathway by interacting with KEAP1, which inactivates the BCR(KEAP1) complex, resulting in nuclear accumulation of NFE2L2/NRF2 and expression of cytoprotective genes. This supplementary information is collated from multiple sources and compiled automatically.

Alternative names

ORCA, OSIL, SQSTM1, Sequestosome-1, EBI3-associated protein of 60 kDa, Phosphotyrosine-independent ligand for the Lck SH2 domain of 62 kDa, Ubiquitin-binding protein p62, EBIAP, p60, p62

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: PBS
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Carrier free: Yes
Clone number: EPR18351
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: +4°C
Storage information: Do Not Freeze

Notes

ab227992 is the carrier-free version of Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab207305.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Compatibility
This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

13 product images

Immunoprecipitation - Anti-SQSTM1 / p62 antibody [EPR18351] - BSA and Azide free (ab227992)
SQSTM1 / p62 was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab207305 at 1/40 dilution.
Western blot was performed from the immunoprecipitate using Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab207305 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/10000 dilution.
Lane 1: HeLa whole cell lysate 10ug (Input).
Lane 2: Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab207305 IP in HeLa whole cell lysate.
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab207305 in HeLa whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 1 second.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab207305).
All lanes: Immunoprecipitation - Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker (Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab207305)
Predicted band size: 47 kDa
Flow Cytometry (Intracellular) - Anti-SQSTM1 / p62 antibody [EPR18351] - BSA and Azide free (ab227992)
SQSTM1 / p62 Flow Cytometry (Intracellular) staining using rabbit Anti-SQSTM1 / p62 antibody
Clone EPR18351 (ab227992) has been successfully conjugated by Abcam. This image was generated using Anti-SQSTM1 / p62 antibody [EPR18351] (PE). Please refer to PE Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab225454 for protocol details.
Overlay histogram showing HeLa cells untreated (red line) and HeLa cells treated with Chloroquine, 50μM, 24 hours, (green line) stained with PE Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab225454. The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (PE Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab225454, 1/5000 dilution) for 30 min at 22°C.
Isotype control antibody (black line) was Rabbit IgG (monoclonal) Phycoerythrin (PE Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab209478) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.
Acquisition of >5,000 events were collected using a 50 mW Yellow/Green laser (561nm) and 586/15 bandpass filter.
This antibody gave a positive signal in HeLa +/- Chloroquine cells fixed with 4% formaldehyde (10 min)/permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions.
Immunocytochemistry/ Immunofluorescence - Anti-SQSTM1 / p62 antibody [EPR18351] - BSA and Azide free (ab227992)
Clone EPR18351 (ab227992) has been successfully conjugated by Abcam. This image was generated using Anti-SQSTM1 / p62 antibody [EPR18351] (Alexa Fluor® 647). Please refer to Alexa Fluor® 647 Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab225453 for protocol details.
Alexa Fluor® 647 Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab225453 staining SQSTM1 / p62 in wild-type HAP1 cells and knockout cells, untreated and chloroquine-treated (50μM, 24 hours). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with Alexa Fluor® 647 Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab225453 at 5μg/ml (shown in red) and Alexa Fluor® 488 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 488), at 1/250 dilution (shown in green) overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Immunocytochemistry/ Immunofluorescence - Anti-SQSTM1 / p62 antibody [EPR18351] - BSA and Azide free (ab227992)
This data was developed using the same antibody clone in a different buffer formulation (Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab207305). Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab207305 was shown to react with SQSTM1 in wild-type U-2 OS cells in Immunocytochemistry with loss of signal observed in a SQSTM1 knockout cell line. Wild-type and knockout cells were mixed and pelleted at a 1:1 ratio on coverslips. The cells were fixed with 4% paraformaldehyde (15 min) then permeabilized with 0.1% Triton X-100 (10min) and then blocked with 1/5000. The cells were then incubated with Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab207305 at 1/200 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat anti-rabbit secondary antibody to (Alexa Fluor^® 555) at 0.5 μg/ml. Acquisition of the green (wild-type), red (antibody staining) and far-red (knockout) channels was performed. Representative grayscale images of the red channel are shown. Wild-type and knockout cells are outlined with yellow and magenta dashed line, respectively. Schematic representation of the mosaic strategy used is shown on the bottom-right panel. Image was acquired with a Zeiss(LSM-880). These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
Western blot - Anti-SQSTM1 / p62 antibody [EPR18351] - BSA and Azide free (ab227992)
SQSTM1 / p62 Western blot staining using rabbit Anti-SQSTM1 / p62 antibody
This data was developed using Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab207305, the same antibody clone in a different buffer formulation.
Lanes 1-4: Merged signal (red and green). Green - Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab207305 observed at 55 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa.
Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab207305 Anti-SQSTM1 / p62 antibody [EPR18351] was shown to specifically react with SQSTM1 / p62 in wild-type HCT116 cells. The band observed in CRISPR/Cas9 edited cell line Human SQSTM1 (p62) knockout HCT116 cell line ab266871 (CRISPR/Cas9 edited cell lysate Human SQSTM1 (p62) knockout HCT116 cell lysate ab257052) lane below 55 kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type and SQSTM1 / p62 CRISPR/Cas9 edited samples were subjected to SDS-PAGE. Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab207305 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (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-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker (Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab207305)
Lane 1: Wild-type HCT116 cell lysate at 20 µg
Lane 2: SQSTM1 CRISPR/Cas9 edited HCT116 cell lysate at 20 µg
Lane 2: Western blot - Human SQSTM1 (p62) knockout HCT116 cell line (Human SQSTM1 (p62) knockout HCT116 cell line ab266871)
Lane 3: HepG2 cell lysate at 20 µg
Lane 4: HeLa cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/10000 dilution
Predicted band size: 47 kDa
Observed band size: 55 kDa
Immunocytochemistry/ Immunofluorescence - Anti-SQSTM1 / p62 antibody [EPR18351] - BSA and Azide free (ab227992)
This ICC data was generated using the same anti-SQSTM1/p62 antibody clone, EPR18351, in a different buffer formulation (cat# Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab207305).
Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab207305 staining SQSTM1 in wild-type HAP1 cells and knockout cells, untreated and chloroquine-treated (50μM, 24 hours). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab207305 at 1μg/ml and Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 at 1/250 dilution (shown in pseudocolour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Western blot - Anti-SQSTM1 / p62 antibody [EPR18351] - BSA and Azide free (ab227992)
SQSTM1 / p62 Western blot staining using rabbit Anti-SQSTM1 / p62 antibody
This data was developed using the same antibody in a different buffer formulation (Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab207305).
Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab207305 was shown to react with SQSTM1 in wild-type U-2 OS cells in Western blot with loss of signal observed in a SQSTM1 knockout cell line. Wild-type U-2 OS and SQSTM1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab207305 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with goat anti-rabbit HRP secondary antibodies at 0.2ug/mL before imaging. These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
All lanes: Western blot - Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker (Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab207305) at 1/1000 dilution
Lane 1: Wild-type U-2 OS cell lysate at 20 µg
Lane 2: SQSTM1 knockout U-2 OS cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 47 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SQSTM1 / p62 antibody [EPR18351] - BSA and Azide free (ab227992)
SQSTM1 / p62 Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining using rabbit Anti-SQSTM1 / p62 antibody
This IHC data was generated using the same anti-SQSTM1/p62 antibody clone, EPR18351, in a different buffer formulation (cat# Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab207305).
Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma tissue labeling SQSTM1 / p62 with Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab207305 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution. Cytoplasm staining on human hepatocellular carcinoma is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-SQSTM1 / p62 antibody [EPR18351] - BSA and Azide free (ab227992)
Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab207305 staining SQSTM1/p62 in HeLa cells +/- Chloroquine (50μM, 24 hours). The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab207305 at 1μg/ml and Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at 1/250 dilution (shown in pseudocolor red) followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab207305).
Immunoprecipitation - Anti-SQSTM1 / p62 antibody [EPR18351] - BSA and Azide free (ab227992)
SQSTM1 / p62 was immunoprecipitated from 1mg of HepG2 (Human liver hepatocellular carcinoma) whole cell lysate with Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab207305 at 1/40 dilution.
Western blot was performed from the immunoprecipitate using Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab207305 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/10000 dilution.
Lane 1: HepG2 whole cell lysate 10ug (Input).
Lane 2: Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab207305 IP in HepG2 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab207305 in HepG2 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 1 second.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab207305).
All lanes: Immunoprecipitation - Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker (Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab207305)
Predicted band size: 32 kDa, 47 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SQSTM1 / p62 antibody [EPR18351] - BSA and Azide free (ab227992)
Immunohistochemical analysis of paraffin-embedded human lung cancer tissue labeling SQSTM1 / p62 with Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab207305 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution. Cytoplasm and weakly nucleus staining on human lung cancer is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab207305).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SQSTM1 / p62 antibody [EPR18351] - BSA and Azide free (ab227992)
Immunohistochemical analysis of paraffin-embedded human liver tissue labeling SQSTM1 / p62 with Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab207305 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution. Negative staining on Hhman normal liver tissue. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab207305).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cytometry (Intracellular) - Anti-SQSTM1 / p62 antibody [EPR18351] - BSA and Azide free (ab227992)
SQSTM1 / p62 Flow Cytometry (Intracellular) staining using rabbit Anti-SQSTM1 / p62 antibody
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed PC-3 (Human prostate adenocarcinoma cell line) cells labeling SQSTM1 / p62 with Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab207305 at 1/100 dilution (red) compared with a rabbit monoclonal IgG isotype control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat Anti-Rabbit IgG (FITC) at 1/500 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 (Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab207305).